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35 publications mentioning dme-mir-34

Open access articles that are associated with the species Drosophila melanogaster and mention the gene name mir-34. Click the [+] symbols to view sentences that include the gene name, or the word cloud on the right for a summary.

1
[+] score: 554
While the magnitude of changes in Diptericin expression upon silencing of individual genes appears to be weaker than that elicited by miR-34 overexpression, possibly due to low knockdown efficiency and/or slow turnover of target proteins, these data nonetheless suggest that CG8468, dlg1 and mura are candidate miR-34 target genes that could potentially downregulate IMD signaling. [score:13]
As most miRNAs only mildly reduce the expression of cognate target genes, we undertook an inclusive approach and considered all mRNAs that display a decrease in expression upon miR-34 over -expression. [score:8]
Importantly, while flies of both genotypes show a decrease in survival following Ecc15 infection, miR-34 over -expression flies display a significantly higher survival rate than control flies (Fig 1I), consistent with the observed higher levels of AMP expression in miR-34 over -expression flies. [score:7]
We show that 1) Dlg1 depletion resembles the phenotype elicited by miR-34 over -expression; 2) miR-34 over -expression causes a marked reduction in both mRNA and protein levels of Dlg1; 3) the 3’ UTR of the dlg1 mRNA contains a functional miR-34-responsive site; and 4) Over -expression of dlg1 abrogates the immune-stimulatory activity of miR-34. [score:7]
A Venn diagram shows the number of candidate miR-34 target genes predicted by TargetScan or PicTar, as well as the number of genes that display a decrease in mRNA levels upon miR-34 over -expression in S2 cell. [score:7]
Identification of miR-34 target genes relevant to innate immunity signalingTo identify miR-34 targets, we first employed two wi dely used bioinformatics algorithms (TargetScan and PicTar) [46, 68, 69]. [score:7]
Notably, this age -dependent increase in basal levels of AMP expression persists in miR-34 [KO] animals, suggesting that age -dependent increase in miR-34 expression does not significantly contribute to the observed difference in AMP expression between young and old flies. [score:7]
In subsequent studies, we focused on miR-34 because 1) miR-34 over -expression causes a concordant increase in levels the Diptericin mRNA in both uninfected and E. coli-infected flies; and 2) miR-34 expression is regulated by ecdysone signaling, which impacts innate immunity [33, 63]. [score:6]
It is possible that miR-34 overexpression may impact ecdysone -mediated regulation of PGRP-LC expression [33], thereby affecting IMD signaling. [score:6]
In addition, with respect to miRNAs, ecdysone treatment markedly activates and inhibits, respectively, the expression of the miRNA genes let-7 and miR-34 [63]. [score:5]
Take together, our study identifies miR-34 as a modulator of innate immunity, identifies both cis-regulatory elements and trans-acting transcription factors required for ecdysone -mediated repression of miR-34, and reveals that the cross-regulation between ecdysone signaling and miR-34 expression contributes to optimal levels of immune activation upon microbial challenge. [score:5]
List of 27 candidate miR-34 target genes identified by TargetScan, PicTar and RNA-sequencing. [score:5]
Lastly, since miR-34 expression steadily increases with age, we also examined AMP expression, host survival and pathogen clearance upon Ecc15 infection in various groups of age-matched miR-34 [KO] and control flies. [score:5]
S4 FigOver -expression of miR-34 in S2 cells activates pirk expression. [score:5]
To identify miR-34 targets, we first employed two wi dely used bioinformatics algorithms (TargetScan and PicTar) [46, 68, 69]. [score:5]
We confirmed the inhibitory effect of 20-HE on miR-34 expression by performing Northern blot to quantify levels of mature miR-34 in S2 cells before and after 20-HE treatment (Fig 5A and 5B). [score:5]
S2 TableList of 27 candidate miR-34 target genes identified by TargetScan, PicTar and RNA-sequencing. [score:5]
We conclude that miR-34 over -expression enhances innate immunity and host survival upon Gram -negative bacterial infection, at least in part, by promoting AMP expression and pathogen clearance. [score:5]
In particular, miR-34 over -expression in cultured cells or in vivo causes aberrant activation of IMD signaling both in the absence and in the presence of immune challenge, and flies over -expressing miR-34 display improved survival and pathogen clearance upon Gram -negative bacterial infection. [score:5]
Importantly, over -expression of the Eip75B-RC isoform, which lacks site #1, significantly suppressed the immune-activation function of miR-34 (Fig 4G). [score:5]
These genes display discrete expression profiles during development and immune activation, as they are likely subject to additional regulatory mechanisms besides miR-34. [score:5]
Next, we examined the impact of miR-34 over -expression on AMP expression in immuno-competent cultured S2 cells. [score:5]
Lastly, we found that while miR-34 over -expression led to a significant reduction in levels of Dlg1 protein, expression of a dlg1 cDNA construct containing only the open reading frame efficiently restored Dlg1 protein levels in S2 cells even in the presence of exogenous miR-34 (Fig 3K, lower panel). [score:5]
Among our list of miR-34 targets is Eip74EF, which encodes a key component of the ecdysone signaling cascade and has been previously identified as a miR-34 target gene [64]. [score:5]
In particular, we show that miR-34 achieves its immune-modulating function, at least in part, by repressing the expression of two novel target genes (Dlg1 and Eip75B). [score:5]
Inspired by the notion that a single miRNA can coordinately regulate the expression of multiple components of a given pathway [71], we performed immunoblot assay and surveyed additional ecdysone response genes to search for additional miR-34 targets. [score:5]
Our analysis revealed that while infected flies of both genotypes carry comparable amount of Ecc15 at the starting time point, miR-34 over -expression flies appear to out-perform control animals in clearing invading pathogens, as indicated by the lower pathogen load in miR-34 over -expression flies than in controls (Fig 1J). [score:5]
Furthermore, our analysis reveals that ecdysone strongly inhibits miR-34 expression via transcriptional repression in a manner that is dependent on a number of transcription factors, including the ecdysone receptor and the Broad Complex (BrC), key mediators of ecdysone signaling cascade. [score:5]
Over -expression of miR-34 in S2 cells impact AMP gene expression. [score:5]
Over -expression of miR-34 in S2 cells activates pirk expression. [score:5]
S3 FigOver -expression of miR-34 in S2 cells impact AMP gene expression. [score:5]
Consistent with the notion that dlg1 is a miR-34 target gene, we observed a reduction in both mRNA and protein levels of dlg1 upon over -expression of miR-34 (Fig 3E and 3F). [score:5]
Over -expression of miR-34 leads to hyperactivation of innate immunity signaling both in cultured cells and in vivo To further reveal the identities of cellular miRNAs that underlie the innate immunity phenotype, we conducted a miRNA over -expression screen in vivo using 101 UAS-miRNA transgenic lines and the da-Gal4; tub-Gal80 [ts] composite line [62]. [score:5]
For miR-34 overexpression experiments, cells were first transfected with a miR-34 expression construct controlled by the metallothionein promoter. [score:5]
We show that over -expression of miR-34 either in flies or in cultured S2 cells leads to hyper-activation of antimicrobial peptide gene expression both in the absence and in the presence of immune challenge, and enhances pathogen clearance in vivo, and that miR-34 deficiency compromises innate immunity. [score:5]
Taken together, these analyses suggest that the regulation of dlg1 gene expression is mediated by both miR-34 -dependent and miR-34-independent mechanisms. [score:4]
Expression of miR-34 is subject to complex regulation during the fly life cycle. [score:4]
Thus, our study lends strong support to the notion that certain miRNAs can coordinately target multiple components of a given biological process to achieve effective regulation by adding miR-34 to such collection of miRNAs [71]. [score:4]
Lastly, the newly identified miR-34 target genes further expand the repertoire of potential negative regulators of IMD signaling. [score:4]
Alternatively, miR-34 may effect IMD signaling by repressing the afore-mentioned cohort of target genes, including dlg1, Eip75B and Su(Z)12, which encode negative regulators of IMD signaling. [score:4]
In addition, levels of the pirk mRNA, which is transcriptionally activated by IMD signaling and encodes a negative regulator of IMD signaling, were also increased both in untreated and PGN -treated cells upon miR-34 over -expression (S4 Fig). [score:4]
Interestingly, our analysis revealed that Drosomycin expression in response to M. luteus infection was significantly decreased in miR-34 over -expressing flies compared with controls (S2 Fig), suggesting that miR-34 differentially impacts IMD and Toll signaling. [score:4]
To generate mutations in the candidate miR-34 -binding site, the afore-mentioned reporter construct was subject to site-directed mutagenesis using Phusion Site-Directed Mutagenesis Kit (Thermo Fisher) according to the manufacturer’s instructions. [score:4]
Further supporting this notion, a miR-34 transgene driven by the metallothionein promoter was robustly expressed in the presence of 20-HE, while its endogenous counterpart under the control of cognate regulatory elements was strongly repressed under the same conditions (Fig 1F). [score:4]
In addition, we performed mRNA sequencing and compared the mRNA expression profiles in cells over -expressing miR-34 with that in control samples. [score:4]
Thus, our study suggests that the cross-regulation between ecdysone signaling and miR-34 expression appears to constitute a positive feedback loop, which may facilitate to achieve appropriate levels of output from ecdysone signaling under various physiological conditions. [score:4]
miR-34 is expressed at relatively low levels during early stages of development and is strongly repressed by ecdysone signaling at larval and pupal stages. [score:4]
In flies lacking miR-34, dys-regulated expression of Eip74EF is linked to accelerated aging in the brain and shortened lifespan [64]. [score:4]
In fact, flies lacking miR-34 develop normally, consistent with the observed low levels of miR-34 expression during early stages of development [64]. [score:4]
Lastly, we identify ecdysone-responsive regulatory elements required for ecdysone -mediated repression of miR-34 expression. [score:4]
For miR-34 target site reporter assays, 150 ng of the Renilla luciferase reporter gene containing wild type or mutant miR-34 -binding site, 25 ng of a firefly luciferase reporter construct [84], and 600 ng of either the miR-34 expression construct or the empty vector were transfected into these cells. [score:4]
miR-34 overexpression (ox) and knockout (ko) flies were injected with a suspension of pHrodo Red E. coli BioParticles Conjugate in PBS. [score:4]
Over -expression of miR-34 leads to hyperactivation of innate immunity signaling both in cultured cells and in vivo. [score:3]
Thus it appears that in adult flies, miR-34 expression prevails and further represses components of ecdysone signaling. [score:3]
miR-34 overexpression or deficiency do not significantly impact bacterial phagocytosis. [score:3]
Furthermore, to examine whether miR-34 over -expression affects hemocyte -mediated bacterial phagocytosis, which is a key mechanism of the cellular immune response, we injected into flies E. coli bio-particles conjugated with a PH-sensitive dye, which becomes fluorescent only after being engulfed by hemocytes and sorted into the acidic endosomal compartment. [score:3]
Nonetheless, under these conditions, lower levels of Diptericin expression was detected in miR-34 [KO] animals than in control animals (Fig 2C and 2D). [score:3]
Over -expression of miR-34 dampens the Toll innate immunity signaling pathway. [score:3]
S14 Fig Suz12 is another miR-34 target gene relevant to innate immunity signaling. [score:3]
Eip75B is another miR-34 target gene that modulates innate immunity signaling. [score:3]
Beside Eip75B, our study also identifies dlg1 as another miR-34 target gene relevant to innate immunity signaling. [score:3]
Flies expressing sh-gfp (ctr) and ko flies carrying a miR-34 rescue construct (Res), respectively, serve as controls. [score:3]
We conclude that besides dlg1, Eip75B is another miR-34 target gene relevant to innate immunity signaling. [score:3]
Expression construct for miR-34: a ~500 bp DNA fragment encoding pri-miR-34 was amplified by PCR and cloned into pRmHa-3 using EcoRI and BamHI restriction sites. [score:3]
miR-34 impacts innate immunity signaling in vivo To further define the role of miR-34 in innate immunity signaling in vivo, we injected either sterile PBS or a suspension of a concentrated culture of the fly pathogen Erwinia carotovora carotovora strain 15 (Ecc15) into miR-34 over -expression or control flies, and monitored fly survival at different time points post injection. [score:3]
S6 FigIdentification of candidate miR-34 target genes. [score:3]
Here we identify Eip75B, another early response gene, as a new target of miR-34. [score:3]
Lastly, our study reveals that ecdysone signaling represses miR-34 by transcriptional inhibition. [score:3]
miR-34 is transcriptionally repressed by ecdysone signalingIt has been reported that 20- hydroxy ecdysone (20-HE) treatment in S2 cells can profoundly alter the expression levels of a number of miRNAs. [score:3]
A S2 cell line was established that stably expresses miR-34 under the control of the copper-inducible metallothionein promoter. [score:3]
Flies over -expressing miR-34 were infected by M. luteus via septic injury. [score:3]
To further define the role of miR-34 in innate immunity signaling in vivo, we injected either sterile PBS or a suspension of a concentrated culture of the fly pathogen Erwinia carotovora carotovora strain 15 (Ecc15) into miR-34 over -expression or control flies, and monitored fly survival at different time points post injection. [score:3]
We found that RNAi -mediated silencing of Su(Z)12, another predicted miR-34 target gene [83], led to an elevation of Diptericin mRNA levels (S14 Fig). [score:3]
1006034.g001 Fig 1Over -expression of miR-34 activates innate immunity signaling. [score:3]
Validating dlg1 and Eip75B as bona fide miR-34 target genes that modulate innate immunity signaling. [score:3]
This analysis revealed that protein levels of Eip75B, a transcriptional factor and component of ecdysone signaling, are significantly reduced by miR-34 over -expression (Fig 4A and 4B). [score:3]
1006034.g004 Fig 4 Eip75B is another miR-34 target gene that modulates innate immunity signaling. [score:3]
In addition, our study reveals a mutual repression between miR-34 expression and ecdysone signaling, and identifies miR-34 as a crucial node in the intricate interplay between ecdysone signaling and innate immunity. [score:3]
S2 FigOver -expression of miR-34 dampens the Toll innate immunity signaling pathway. [score:3]
This analysis revealed that miR-34 [KO] flies express significantly lower levels of the Diptericin mRNA than control animals, under both non-infection and E. coli infection conditions (Fig 2B). [score:3]
Taken together, these data demonstrate that dlg1 is a bona fide (perhaps a major) miR-34 target gene relevant to IMD signaling. [score:3]
Furthermore, our bioinformatics analysis predicted a miR-34 binding site in the dlg1 3’ un translated region (3’ UTR) (Fig 3I). [score:3]
Over -expression of miR-34 activates innate immunity signaling. [score:3]
Validating dlg1 and Eip75B as bona fide miR-34 target genes that modulate innate immunity signalingDlg1 is a member of membrane- associated guanylate kinase (MAGUK) family proteins (albeit it lacks catalytic activity), and is localized to septate junctions [70]. [score:3]
In addition, levels of the BrC protein, which is a transcriptional factor and component of ecdysone signaling, is also significantly reduced upon miR-34 over -expression (Fig 4A and 4B). [score:3]
Notably, the genomic fragment F3, which displays the highest degree of BrC occupancy, centers around nucleotides 573–587 (Fig 6E and 6F), suggesting that BrC binding at this region is required, at least in part, for the repressive effect of ecdysone on miR-34 expression. [score:3]
Also shown are fold changes in the corresponding mRNA levels upon miR-34 overexpression. [score:3]
Thus, it appears that miR-34 modulates innate immunity signaling by repressing multiple target genes. [score:3]
The AMP expression phenotype for select miRNAs (miR-34, miR-92a, miR-9a and miR-989) is shown (Fig 1D and 1E). [score:3]
Importantly, expression of dlg1 efficiently blunted the immune-stimulating effect of miR-34 (Fig 3K, upper panel). [score:3]
To further examine whether enhanced survival of miR-34 over -expression flies upon Ecc15 infection is attributable to improved pathogen clearance or immune tolerance, we monitored pathogen load at various time points post Ecc15 infection. [score:3]
We show that Dlg1 and Eip75B are two novel miR-34 target genes relevant to innate immunity. [score:3]
RNA-Seq datasets containing ~25 million 2 x 90 bp reads were generated from control S2 cells or cells over -expressing miR-34 using an Illumina HiSeq 2000 system. [score:3]
To alleviate the concern that endogenous or environmental microbes could potentially impact host innate immunity and cause variations in levels of AMP expression, we also analyzed miR-34 [KO] and wildtype flies raised in media containing multiple antibiotics. [score:3]
Lastly, we show that miR-34 represses the expression of a number of components in ecdysone signaling, including Eip74EF, Eip75B and BrC. [score:3]
S2 cells stably expressing miR-34 were generated by transfection with pRmHa-3- miR-34 and the selection marker plasmid pHS-neo using the calcium phosphate method, followed by selection in medium containing 400 μg/mL G418 (Calbiochem). [score:3]
Considering previous reports showing that innate immunity signaling is activated under various stress conditions and in aged flies, and that levels of Diptericin display an increase during aging [65– 67], it is possible that elevated miR-34 expression in aged flies may contribute to an increase in stress, which in turn leads to activation of innate immunity signaling pathways. [score:3]
In addition, depletion of Eip75B in S2 cells resembles the miR-34 over -expression phenotype, i. e. activation of innate immunity signaling both in the presence and absence of PGN treatment (Fig 4E and 4F). [score:3]
Identification of miR-34 target genes relevant to innate immunity signaling. [score:3]
Such ecdysone -mediated modulation of miR-34 gene expression is most likely at the level of transcription, as changes in levels of the primary miRNA transcripts resemble that of mature miRNAs (Fig 5A–5C) [63]. [score:3]
Taken together, these data demonstrate that mis -expression of miR-34 affects the IMD innate immunity signaling pathway both in cultured cells and in vivo. [score:3]
This analysis reveals a similar degree of phagocytosis between control and miR-34 over -expression flies (S5A and S5B Fig). [score:3]
These data demonstrate that miR-34 deficiency compromises AMP expression and impairs IMD signaling. [score:3]
dlg1 is a miR-34 target gene relevant to innate immunity signaling. [score:3]
This analysis revealed that the repressive effect of ecdysone on miR-34 expression was partially relieved in EcR or BrC knockdown cells compared with control RNAi cells (Fig 5A–5F). [score:3]
Integration of these datasets identified a list of 27 genes that not only scored positively using both bioinformatics algorithms, but also displayed a decrease in mRNA levels upon miR-34 over -expression (S6 Fig & S2 Table). [score:3]
1006034.g003 Fig 3 dlg1 is a miR-34 target gene relevant to innate immunity signaling. [score:3]
However, activation of IMD signaling does not appear to strongly affect miR-34 expression, as levels of miR-34 remain essentially unchanged in the body of flies upon E. coli infection (S11B and S11C Fig). [score:3]
This is consistent with the notion that a functional miR-34 target site is present only in the 3’ UTR of the Dlg1 mRNA. [score:3]
Interestingly, in ecdysone -treated S2 cells the effect of miR-34 overexpression appears to be PGRP-LC -dependent, even in the absence of PGN stimulation. [score:3]
Eip74EF, a validated miR-34 target gene, is among such group of early response genes. [score:3]
Suz12 is another miR-34 target gene relevant to innate immunity signaling. [score:3]
Furthermore, consistent with the findings of Silverman, Ambros and colleagues [33, 63], we detected higher levels of pri-miR-34 in BrC mutant larvae than in control animals, and found that the BrC mutant larvae expressed lower levels of Dpt in response to E. coli infection (Fig 5G and 5H). [score:3]
S5 Fig miR-34 overexpression or deficiency do not significantly impact bacterial phagocytosis. [score:3]
We confirmed these findings by showing that over -expression of miR-34 can lead to a significant reduction in the levels of Eip74EF protein (Fig 4A and 4B). [score:3]
We detected higher levels of the Diptericin mRNA, as well as a panel of mRNAs encoding additional AMPs, including Cecropin A1 (CecA1), Attacin A (AttA), Metchnikowin (Mtk) and Defensin (Def), in miR-34 overexpressing cells than in control cells, both in the absence and presence of peptidoglycan (PGN) treatment (Fig 1F and 1G, S3 Fig). [score:3]
Identification of candidate miR-34 target genes. [score:3]
In addition, miR-34 modulates IMD signaling, in part, by repressing genes encoding the septate junction protein Dlg1 as well as Eip75B, a component of the ecdysone signaling cascade and a negative regulator of the IMD pathway. [score:2]
In addition, higher levels of Dlg1 protein were detected in miR-34 knock out flies than in control animals (Fig 3G and 3H). [score:2]
These observations suggest that BrC may directly contribute to ecdysone -mediated repression of miR-34 by binding to the F3 region. [score:2]
To identify the regulatory elements required for ecdysone -mediated repression of miR-34, 100 ng of miR-34 enhancer- firefly luciferase plasmid and 100 ng of pol III-Renilla luciferase or 20 ng of actin-Renilla luciferase were transfected into cells. [score:2]
Importantly, in both young and aging settings, miR-34 [KO] flies display lower basal and E. coli infection -induced levels of Diptericin expression, poorer survival and pathogen clearance in response to Ecc15 challenge compared to age-matched control animals (Fig 2C–2H). [score:2]
Identification of cis-regulatory elements responsible for ecdysone -mediated repression of miR-34 The miR-34 genomic locus contains two additional miRNA genes, miR-277 and miR-317, as well as a protein-coding gene Fmr1 (Fig 6A). [score:2]
In addition, we generated a corresponding set of reporter constructs carrying mutations in candidate miR-34 binding sites that abolish miR-34 recognition. [score:2]
Taken together, these analyses identified key cis-regulatory genomic elements and trans-acting transcription factors required for ecdysone -mediated repression of miR-34. [score:2]
Mapping cis-regulatory elements required for ecdysone -mediated repression of miR-34. [score:2]
We identify a cis-regulatory region (encompassing F3, Fig 6E) from the miR-34 locus required for optimal ecdysone -mediated repression of miR-34. [score:2]
To further define the minimal cis-regulatory element within peak P2 that are required for miR-34 repression, we placed various truncated P2 genomic fragment upstream of the reporter gene and assessed their capability of conferring ecdysone -mediated silencing. [score:2]
This analysis reveals that miR-34 is capable of silencing the reporter construct carrying a wildtype dlg1 3’ UTR, and this repression is relieved by introducing mutations in the miR-34 binding site (Fig 3J). [score:2]
Identification of cis-regulatory elements responsible for ecdysone -mediated repression of miR-34. [score:2]
Lastly, we identify key cis-regulatory genomic elements and trans-acting transcription factors required for optimal ecdysone -mediated repression of miR-34. [score:2]
1006034.g006 Fig 6Mapping cis-regulatory elements required for ecdysone -mediated repression of miR-34. [score:2]
Thus our study uncovers miR-34 as a component of an ecdysone -dependent regulatory circuit that modulates IMD innate immunity signaling in Drosophila. [score:2]
Next, we examined whether miR-34 deficiency impacts innate immunity by measuring AMP expression both in miR-34 knockout (miR-34 [KO]) flies [64] and in control (Res) animals (miR-34 [KO] flies carrying a miR-34 genomic rescue construct) (Fig 2A). [score:2]
Here we show that the conserved miRNA miR-34 regulates antibacterial defense and steroid hormone signaling in Drosophila. [score:2]
Consistent with lower levels of Dpt expression, miR-34 [KO] flies present poorer survival and pathogen clearance in response to Ecc15 compared to control animals (Fig 2E). [score:2]
In this study, we identify the microRNA miR-34 as a link in the intricate interplay between ecdysone signaling and innate immunity. [score:1]
Among these 27 genes is Eip74EF, which is a well-characterized miR-34 target, thereby validating our approach [64]. [score:1]
In addition, considering that the miR-34 gene itself is transcriptionally repressed by ecdysone signaling, our findings indicate that ecdysone signaling not only activates select EIP genes transcriptionally to boost levels of the corresponding mRNAs, but also maximize the protein output from these mRNAs by reducing levels of miR-34. [score:1]
In contrast, miR-34 mutant flies present profound defects in antibacterial innate immunity. [score:1]
miR-34 impacts innate immunity signaling in vivo. [score:1]
Of note, since the miR-34 effect on immunity is PGRP-LC -dependent, it would be interesting to assess potential interactions between Dlg1 and PGRP-LC. [score:1]
1006034.g005 Fig 5Identification of trans-acting transcription factors required for ecdysone -mediated repression of miR-34. [score:1]
1006034.g002 Fig 2 miR-34 deficiency compromises innate immunity. [score:1]
We noticed that while BrC dsRNA treatment led to a significant reduction in both mRNA and protein levels of BrC, 20-HE -mediated repression of miR-34 was only partially relieved (Fig 5A–5F). [score:1]
The miR-34 genomic locus contains two additional miRNA genes, miR-277 and miR-317, as well as a protein-coding gene Fmr1 (Fig 6A). [score:1]
Of note, levels of miR-34 remain unchanged in E. coli-infected flies (S11B and S11C Fig), indicating that the observed decrease in Dlg1 protein levels is attributable to a miR-34-independent mechanism. [score:1]
Our analysis revealed that three additional transcription factors (Srp, Twi and Ap) are required for optimal repression of miR-34 by ecdysone (Fig 5I, S9 Fig). [score:1]
Fly survival was recorded daily and plotted (n≥3; p<0.001 between Enterobacter cloacae-infected control and miR-34 [KO] flies). [score:1]
1006034.g007 Fig 7A schematic summary of the role of miR-34 in modulating IMD innate immunity signaling. [score:1]
In contrast, levels of miR-34 display a significant decrease in response to 20-HE treatment. [score:1]
These observations suggest that there are yet-to-be-identified factors that are required for 20-HE -mediated miR-34 repression. [score:1]
Interestingly, the identified genomic element coincides with the region that displays a high degree of occupancy by BrC, an ecdysone -induced transcription factor required for the optimal repression of miR-34 by ecdysone. [score:1]
In fact, miR-34 knockout flies display an early onset of neuro-degeneration during aging compared with age-matched wildtype counterparts. [score:1]
Taken together, these data demonstrate that both EcR and BrC are required for optimal ecdysone -mediated transcriptional repression of miR-34. [score:1]
Considering a previous report showing that let-7 represses Diptericin [59] and the pro-immunity role of miR-34 uncovered in this study, it would have been expected that ecdysone would reduce the output of the IMD innate immunity signaling pathway. [score:1]
Reporter constructs containing candidate miR-34 -binding sites identified in the Eip75B ORF: pairs of oligos containing triple copies of individual sites (wildtype or mutant) were annealed and cloned into pRmHa-3-Renilla using BamHI and SalI sites. [score:1]
Besides IMD signaling, which is activated upon Gram -negative bacterial infection, we also asked whether miR-34 additionally impacts other signaling routes, such as the Toll signaling pathway, which mediates host defense against infection by fungi and Gram -positive bacteria. [score:1]
A schematic summary of the role of miR-34 in modulating IMD innate immunity signaling. [score:1]
Seed region of miR-34 was highlighted in green. [score:1]
miR-34 deficiency compromises innate immunity. [score:1]
In addition, while BrC has been implicated in ecdysone -mediated repression of miR-34, the requirement for BrC in this process could not be definitively assessed [63]. [score:1]
miR-34 is transcriptionally repressed by ecdysone signaling. [score:1]
Since BrC is required for optimal ecdysone -mediated repression of miR-34, we inspected the nucleotide sequence of peak P2 and identified five putative BrC -binding sites (shown as shaded boxes in Fig 6E). [score:1]
Strikingly, miR-34 levels are significantly elevated in adult flies upon eclosure and with age [64]. [score:1]
Thus our study identifies miR-34 as a node linking steroid hormone signaling and immunity, thereby enriching the repertoire of immune-modulating miRNAs in animals and providing insights into the interplay between steroid hormone signaling and innate immunity. [score:1]
Of note, the BrC transcript harbors a predicted miR-34 binding site, which could underlie miR-34 -mediated repression [46, 68]. [score:1]
In addition, we found that the immune-modulating role of miR-34 is critically dependent on IMD signaling, and that miR-34 operates in part by repressing genes encoding the septate junction protein Dlg1 and the nuclear hormone family transcription factor Eip75B, a key mediator of the ecdysone steroid hormone signaling cascade. [score:1]
In summary, our study identifies the conserved miRNA miR-34, together with several other miRNAs, as modulators of innate immunity in Drosophila. [score:1]
Lastly, our analysis reveals that miR-34 [KO] flies display a defective survival and pathogen clearance in response to Enterobacter cloacae infection (Fig 2I and 2J), and that miR-34 deficiency did not significantly impact hemocyte -mediated phagocytosis (S5C and S5D Fig). [score:1]
S7 FigThe Eip75B transcript harbors a second miR-34 -binding site. [score:1]
We therefore examined whether these transcription factors contribute to ecdysone -mediated repression of miR-34. [score:1]
Identification of trans-acting transcription factors required for ecdysone -mediated repression of miR-34. [score:1]
Furthermore, our analyses reveal that miR-34 is transcriptionally repressed by the ecdysone signaling cascade in a manner that is dependent on the ecdysone receptor and the transcription factor BrC. [score:1]
Fly survival was recorded daily up to day 8 post-infection and plotted (n≥3; p<0.05 between Ecc15-infected control and miR-34 [OX] (da>miR-34 Gal80 [ts]) flies). [score:1]
This analysis reveals that reporter genes containing wild type sites #1 or #5 were more efficiently repressed by miR-34 than the reporter with mutant sites (Fig 4C and 4D & S7 Fig). [score:1]
This led to the identification of miR-34 among several other miRNAs as modulators of IMD signaling. [score:1]
In contrast, levels of miR-34 increase drastically in newly hatched adults and display a further elevation with age [64]. [score:1]
Importantly, depletion of core components of the IMD signaling pathway, such as PGRP-LC, subunits of the DmIKK complex (Ird5 and Kenny), or Relish, significantly alleviated the immune-activation phenotype of miR-34 (Fig 1H), indicating that the canonical IMD signaling pathway is required for the immune-stimulating function of miR-34. [score:1]
Fly survival was recorded daily and plotted (n≥3; p<0.001 between Ecc15-infected control and miR-34 [KO] flies). [score:1]
In addition, our study reveals a mutual repression between miR-34 and steroid hormone signaling and identifies genomic elements and transcription factors required for steroid hormone -mediated repression of miR-34. [score:1]
Further inspection of the Eip75B mRNA sequence allowed us to identify five potential miR-34 sites in the coding region. [score:1]
This approach has identified several genomic regions near the miR-34 locus (referred to as ecdysone-responsive peaks P1 through P5) as putative ecdysone-responsive elements (Fig 6A) [75]. [score:1]
In order to test whether any of these sites can confer miR-34 -mediated gene silencing, we generated reporter constructs by placing tandem triple repeats of individual miR-34 binding sites derived from the Eip75B mRNA in the 3’ UTR of the firefly luciferase gene, and examined whether these reporter genes can be repressed by miR-34. [score:1]
Furthermore, our findings showing that miR-34 can repress multiple genes of ecdysone signaling, including Eip74EF, Eip75B and BrC, add an additional layer of complexity. [score:1]
The Eip75B transcript harbors a second miR-34 -binding site. [score:1]
Of note, three additional transcription factors, including Srp, Twi and Ap, also contribute to ecdysone -mediated repression of miR-34, and predicted Srp- and Twi -binding sites can be found within and close to F3, respectively (Fig 6E). [score:1]
To generate various miR-34 enhancer- firefly luciferase plasmid, Selected regions were PCR amplified from genomic DNA, cloned into pCR8-TOPO-GW (Invitrogen), and shuttled to pGL3_GW_luc+ [76] by LR clonase II recombination (Invitrogen). [score:1]
miR-34 over-production or deficiency, respectively, enhances or impairs antibacterial defense. [score:1]
As a consequence, ecdysone signaling prevails and keeps miR-34 levels low. [score:1]
Note that due to lethality, 1 fly per group was used for a subset of data points in miR-34 [KO] flies 2 days post-infection. [score:1]
[1 to 20 of 191 sentences]
2
[+] score: 273
Other miRNAs from this paper: dme-mir-1
Because the GAL4-OK107 expresses GAL4 in MB progenitor cells, including neuroblasts and ganglion mother cells, as well as the MB γ, α′/β′ and α/β neuronal types 21 22, we speculated that the effect of ectopically overexpressed miR-34 on γ lobe pruning in MB neurons might be caused by a miR-34 -induced change in the developmental fate of the MB progenitor cells that resulted in the elimination of axon pruning, rather than the direct inhibition of axon pruning in differentiated MB γ neurons. [score:9]
Using an experimental approach in which miR-34 was ectopically expressed, we found that miR-34, which is known to protect against chronic neurodegeneration in the aged Drosophila brain, can inhibit developmentally related axon pruning in MB γ neurons (Fig. 2b), but cannot inhibit axotomy -induced axon degeneration in OSN axons (Supplemental Figure 1). [score:8]
These results collectively suggested that ectopic overexpression of miR-34 results in defective axon pruning in MB γ neurons through the downregulation of EcR-B1 expression. [score:8]
However, we cannot confirm whether babo is a direct target of miR-34 since we observed no obvious change in Babo expression in wild-type, miR-34 overexpression and mir-34 mutant flies based on western blot analysis using an anti-Babo antibody (ab14682; Abcam; Lai, et al., unpublished observation). [score:8]
Ectopic miR-34 overexpression downregulates the endogenous EcR-B1 expression in MB neurons exhibiting defective γ lobe pruning. [score:8]
Ectopic miR-34 overexpression downregulated EcR-B1 expression in MB neurons. [score:8]
Confocal images show the lobe pruning phenotypes of MB neurons in flies with ectopic overexpression of EcR-B1 (a), ectopic overexpression of miR-34 (b), or the overexpression of both (c). [score:7]
Although the results of our experiments suggested that miR-34 influenced γ axon pruning via the downregulation of EcR-B1 expression, it remained unclear whether other miR-34-regulated genes play essential roles in controlling axon pruning in MB γ neurons. [score:7]
We also used Asense-GAL80 in separate experiments to suppress GAL4-OK107 driven miR-34 overexpression in neural progenitors and young neurons, but not in differentiated MB neurons 24, and this manipulation cannot inhibit the miR-34 -induced γ lobe pruning defect (Supplemental Figure 3, Supplemental Table 1). [score:7]
Confocal images show lobe pruning phenotypes of MB neurons in flies with ectopic overexpression of Babo-a (a), ectopic overexpression of miR-34 (b), or the overexpression of both (c). [score:7]
How to cite this article: Lai, Y. -W. et al. Drosophila microRNA-34 Impairs Axon Pruning of Mushroom Body g Neurons by Downregulating the Expression of Ecdysone Receptor. [score:6]
Notably, we also found that overexpression of EcR-B1 or Babo-a, but not the β-galactosidase control, rescued miR-34 -induced defective γ lobe pruning in MB neurons, which suggested that miR-34 may regulate EcR-B1 expression via the Babo/EcR signaling pathway (Figs 5 and 6, Supplemental Table 1; data not shown, n = 20 for the β-galactosidase control). [score:6]
Our investigation showed that miR-34 mediates defects in γ lobe pruning in differentiated MB neurons via the downregulation of EcR-B1 expression in MB γ neurons (Figs 3, 4 and 5), and that the overexpression of Babo-a rescued the miR-34 -induced γ lobe pruning defect in MB neurons (Fig. 6). [score:6]
These results suggested that the miR-34 -induced impairment of γ axon pruning in MB neurons was caused by miR-34 overexpression in differentiated MB neurons, and was not the result of altered developmental fate due to miR-34 overexpression in progenitor cells and young neurons. [score:6]
Because the adult-onset elevation of miR-34 expression plays an essential role in inhibiting chronic neural deterioration in the aged brain 7, we investigated whether miR-34 also functions in the inhibition of axotomy -induced axon degeneration in OSNs or developmentally related γ axon pruning in MB neurons. [score:6]
Using this strategy in differentiated MB γ neurons, we observed that the effect of ectopically overexpressed miR-34 on the defective γ lobe pruning phenotype was dose -dependent, in which one copy of miR-34 exhibited a moderate, less-penetrant phenotype (80%, n = 5; Fig. 3b, Supplemental Table 1) and two copies of miR-34 overexpression exhibited a severe, fully-penetrant phenotype (100%, n = 13; Fig. 3c, Supplemental Table 1). [score:5]
The defective γ lobe pruning phenotype was rescued by overexpression of Babo-a in MB neurons with ectopic miR-34 overexpression (c). [score:5]
Confocal images show EcR-B1 expression in MB neurons in wild-type flies (a, c) and flies for ectopic miR-34 overexpression (one copy of miR-34 in b; two copies of miR-34 in d) using the mosaic analysis with a repressible cell marker (MARCM) system. [score:5]
We were, however, unable to examine the effect of GAL4-201Y -driven miR-34 expression on MB γ lobe pruning beyond the mid-pupal stage due to toxicity related to the ectopic overexpression of miR-34 (data not shown). [score:5]
The defective γ lobe pruning phenotype was rescued by overexpression of EcR-B1 in MB neurons with ectopic miR-34 overexpression (c). [score:5]
We also used a computational strategy to perform a genome-wide search of genes possessing 3' UTRs with miRNA target sites, in which the Eip74EF, Hr4, and yem transcripts were identified as potential miR-34 targets (Supplemental Figure 4a,b). [score:5]
Ectopic miR-34 overexpression inhibited γ lobe pruning in MB neurons. [score:5]
Thus, it remains unclear whether the expression of proteins involved in γ lobe pruning, other than EcR-B1, are also influenced by the level of miR-34 expression. [score:5]
org/), we identified Eip74EF, hormone receptor 4 (Hr4), and yemanuclein (yem) as mRNAs possessing 3′ untranslated regions (UTRs) with miR-34 target sites (Supplemental Figure 4a). [score:5]
On the other hand, using the pan MB neuronal driver, GAL4-OK107 that expresses GAL4 in MB α/β, α′/β′ and γ neurons 21, the ectopic overexpression of miR-34 caused defects in the γ lobe formation in which excessive axonal branches often projected adjacent to the MB α and β lobes (Fig. 1a,b, Supplemental Table 1). [score:5]
The formation of aberrant larval-specific γ lobes was also observed in MB neurons in which miR-34 was ectopically overexpressed using GAL4-201Y (Fig. 2i,l, Supplemental Table 1), which expresses GAL4 in differentiated MB γ neurons and a small subset of core MB α/β neurons 18. [score:5]
Future investigations are warranted to identify and characterize other miR-34 target genes that might contribute to the regulation of EcR-B1 expression in axon pruning in MB γ neurons to obtain further insight into the regulation of developmentally related large-scale axon degeneration. [score:4]
This miR-34 induced lobe defect is similar to that of the γ lobe phenotype observed in MB neurons in which USP expression was knocked down by RNAi (double arrowhead, c). [score:4]
Among these, Eip74EF has previously been shown to be an miR-34 target that functions in the regulation of neurodegeneration 7, and all three genes are annotated as having transcription factor and DNA binding activities. [score:4]
Confocal images show the lobe phenotype of mushroom body (MB) neurons in wild-type flies (a) and flies with ectopic miR-34 overexpression (b) and RNAi knockdown of USP (c). [score:4]
It is also possible that miR-34 regulated the expression of EcR-B1 through pathways that function parallel to Babo-a, such as TGF-β-independent Fushi tarazu transcription factor 1 (Ftz-f1) and Hormone receptor-like in 39 (Hr39) mediated signaling pathways, which have also been shown to contribute to axon pruning in MB γ neurons 29. [score:4]
Future studies are warranted to examine the effects of miR-34 on the regulatory influence of Myo, Plum, Ftz-f1 and Hr39 on EcR-B1 expression. [score:4]
The expression of EcR-B1 was significantly reduced by ectopic miR-34 overexpression in the MARCM clones (b,d), compared to that of the wild-type MARCM clones (a,c). [score:4]
By contrast, in the MB neurons with ectopic miR-34 overexpression, a significant fraction of larval-specific γ lobes was remained at 18–24 h APF (double-arrowheads, e,f,i), and aberrant axonal braches (most likely γ lobe-derived) were located adjacent to the developing α and β lobes at 36-48 h APF (double-arrowheads, j,k). [score:3]
Defective γ lobe pruning was more severe in flies for ectopic overexpression of two copies of miR-34 transgenes in MB γ neurons (double arrowheads, c). [score:3]
It is possible that γ axon pruning in MB neurons might be mediated by a combination of Eip74EF, Hr4, yem, and/or other yet-to-be-identified miR-34 target genes. [score:3]
These results collectively suggested that the overexpression of miR-34 impaired axon pruning in MB γ neurons. [score:3]
Confocal images show axon pruning of MB neurons in wild-type flies (a) and flies with ectopic miR-34 overexpression using the mosaic analysis with a repressible cell marker system (b, c). [score:3]
By contrast, a substantial fraction of larval-specific γ lobes was preserved at 24 h APF in MB neurons in which miR-34 ectopic overexpression was driven by GAL4-OK107 (Fig. 2d–f). [score:3]
Overexpression of EcR-B1 rescued the miR-34 -induced defective γ lobe phenotype in MB neurons. [score:3]
Confocal images show γ lobes of MB neurons in wild-type flies (a–c, g–i) and flies that were for ectopic miR-34 overexpression (one copy of miR-34 in (d–f, j, k); two copies of miR-34 in l) driven by GAL4-OK107 (a–h, j, k) or GAL4-201Y (i and l). [score:3]
Overexpression of Babo-a rescued the miR-34 -induced defective γ lobe phenotype in MB neurons. [score:3]
Because the defective MB γ lobe pruning phenotype was induced by miR-34 overexpression in nearly all differentiated MB neurons, we speculated whether a cell-autonomous mechanism was involved in the defective axon pruning phenotype. [score:3]
Ectopic miR-34 overexpression in differentiated MB neurons disrupted γ axon pruning. [score:3]
Ectopic miR-34 overexpression caused aberrant lobe formation on mushroom body (MB) neurons. [score:3]
To investigate this possibility, we used Asense-GAL4, which expresses GAL4 in neural progenitors and young neurons but not in differentiated neurons 23, to drive the expression of miR-34. [score:3]
Ectopic miR-34 overexpression in MB neurons caused defective γ lobe pruning (double-arrowheads, b). [score:3]
The aberrant larval-specific γ lobes persisted at 36 and 48 h APF in flies with ectopically overexpressed miR-34, but not in wild-type flies (Fig. 2g,h,j,k). [score:3]
Therefore, we investigated whether ectopically expressed miR-34 also reduces the expression of EcR-B1 in MB neurons that in turn elicits the defective γ lobe pruning phenotype. [score:3]
No defect in γ lobe pruning was observed in MB neurons derived from Asense-GAL4 [+] progenitor cells and young neurons in which miR-34 was overexpressed (Supplemental Figure 2, Supplemental Table 1). [score:3]
To test this hypothesis, we applied the MARCM system to ectopically express miR-34 using GAL4-OK107 and GAL4-201Y in MB neurons in separate experiments. [score:3]
These results combined with those of the analysis of stage-specific miR-34 overexpression suggested that the induction of the miR-34 -mediated perturbation of γ axon pruning occurred after the differentiation of MB γ neurons. [score:3]
A subset of the dorsal α lobe was observed on MB neurons in wild-type flies (upper arrow, a), whereas an aberrant axonal bundle (likely unpruned γ lobe) projected outside the MB α lobe (double arrowhead, b) and most of the γ lobes were intact in the sample with ectopic expression of one copy of mir-34 transgene (arrowhead, b). [score:3]
These results collectively suggested that miR-34 overexpression may disrupt axon pruning in MB γ neurons. [score:3]
Ectopic miR-34 overexpression in differentiated MB γ neurons causes aberrant axon pruning. [score:3]
We found that, in wild-type OSNs and those in which miR-34 was ectopically overexpressed, degeneration was induced 3 days following axotomy (Supplemental Figure 1), suggesting that miR-34 does not protect against axotomy -induced axon degeneration in OSNs. [score:3]
Ectopic overexpression of miR-34 impairs the mushroom body (MB) γ lobe pruning. [score:3]
RNAi knockdown of Eip74EF, Hr4, and yem does not affect miR-34 -induced defective γ lobe pruning in MB neurons. [score:2]
We confirmed that the 3′ UTRs of Eip74EF, Hr4, and yem contain functional miR-34 target sites using a previously described luciferase reporter gene assay 26 (Supplemental Figure 4b). [score:2]
The following fly strains used in this study: (1) UAS-mCD8::GFP on the second chromosome 25; (2) Or88a-GAL4 31; (3) GAL4-OK107 21; (4) UAS-mir-34 [[1]]; (5) UAS-mir-34 [[2]]; (6) UAS-usp RNAi 32; (7) hs-FLP [[122]] 33; (8) FRT [[G13]],UAS-mCD8::GFP,GAL4–201Y 18; (9) FRT [[82B]],tubP-GAL80 25; (10) FRT [[82B]] 34; (11) yw,hs-FLP [[1]],UAS-mCD8::GFP 25; (12) UAS-EcR-B1 20; (13) UAS-baba-a 35; (14) UAS-lacZ. [score:1]
miR-1 was transfected as a negative control for the miR-34 experiments. [score:1]
Standard molecular biological techniques were used to generate the UAS-mir-34, UAS-Eip74EF RNAi, and UAS-yem RNAi transgenes. [score:1]
Production of UAS-mir-34, UAS-Eip74EF RNAi, and UAS-yem RNAi transgenic fliesStandard molecular biological techniques were used to generate the UAS-mir-34, UAS-Eip74EF RNAi, and UAS-yem RNAi transgenes. [score:1]
The production of transgenic flies carrying the UAS-mir-34, UAS-Eip74EF RNAi and UAS-yem RNAi transgenes was performed by BestGene (Chino Hills, CA, USA). [score:1]
Production of UAS-mir-34, UAS-Eip74EF RNAi, and UAS-yem RNAi transgenic flies. [score:1]
We removed the antennae and maxillary palps of flies, thereby separating OSN axons from their soma, to establish a system for examining whether miR-34 influences axon degeneration following axotomy, as previously described 10. [score:1]
[1 to 20 of 66 sentences]
3
[+] score: 40
Other miRNAs from this paper: dme-mir-14, dme-mir-9c, dme-mir-989
To determine whether these putative inhibitors of the piRNA pathway are part of a single biological process that antagonizes piRNA -mediated TE repression, we performed a gene ontology (GO) term enrichment analysis on the miR-14 and miR-34 target genes using GOrilla (http://cbl-gorilla. [score:5]
1005194.g007 Fig 7GO term comparisons of miR-14 and miR-34 putative targets and of all the Drosophila miRNA putative targets. [score:5]
GO term comparisons of miR-14 and miR-34 putative targets and of all the Drosophila miRNA putative targets. [score:5]
We compared the 153 miR-14 and the 98 miR-34 putative targets using as background set the 3759 genes that are putatively targeted by all Drosophila miRNAs. [score:4]
org/fly_12/), we found 153 and 98 putative targets for miR-14 and miR-34, respectively. [score:3]
The miR-34 target genes only overlapped modestly with the GO term “basal lamina component”. [score:3]
We observed similar phenotypes upon individual titration (by expression of the corresponding miR-sponge) of miR-14 and miR-34, and also in a miR-14 loss of function mutant. [score:3]
We report here that, in Drosophila somatic ovarian tissues, two miRNAs, miR-14 and miR-34, are required for the accumulation of piRNAs that prevent the expression of transposable elements and, probably, the subsequent invasion of the germinal genome. [score:3]
Next we wanted to identify the gene(s) that are regulated by miR-14 and miR-34 for piRNA -mediated TE repression in follicle cells. [score:2]
Moreover, like upon drosha and AGO1 knock down, the level of two piRNAs (ZAM and Tabor), quantified by RT-qPCR, was clearly decreased following miR-SP -induced miR-14 and miR-34 titration (Fig 6D). [score:2]
For instance, oogenesis did not seem to be affected when miR-sponge -mediated titration of either miR-14 or miR-34 resulted in gypsy- and ZAM-lacZ de-repression. [score:1]
Two Drosha -dependent miRNAs, miR-14 and miR-34, are therefore individually required for both TE repression and TE-derived piRNA accumulation in follicle cells. [score:1]
MiR-SP mediated titration of two miRNAs (miR-14 and miR-34) resulted in lacZ de-repression, as indicated by β-Gal staining and RT-qPCR (Fig 6A– 6B). [score:1]
miR-14 and miR-34 are specifically required for TE piRNA biogenesis and/or stability in follicle cells. [score:1]
Moreover, we report that individual titration of two miRNAs (miR-14 and miR-34) leads to a similar TE de-repression phenotype. [score:1]
[1 to 20 of 15 sentences]
4
[+] score: 24
Other miRNAs from this paper: dme-mir-305, dme-let-7
Expression pattern of Nbr in vivoNibbler is a 3′-to-5′ exonuclease whose function is critical for trimming the 3′end of miR-34-5p in Drosophila (Han et al., 2011; Liu et al., 2011). [score:3]
Importantly, rescue of miRNA trimming activity was dependent on the catalytic residues of the Nbr exonuclease domain: mutating catalytic residues (D435A,E437A) within the genomic transgene (referred to as ‘cat-dead nbr’) failed to rescue miR-34-5p trimming, despite robust protein expression (Fig. 2C). [score:3]
The role of trimming by Nbr on miRNA function has been assessed with the study of miR-34 upon expression of a catalytically mutant form of nbr in cultured cells, where miR-34 silencing was compromised by ∼20% (Han et al., 2011). [score:3]
The genomic rescue construct expressing catalytically dead Nbr failed to rescue the miR-34 isoform pattern, despite robust levels of the Nbr protein. [score:3]
Screening defined a 3′-to-5′ exonuclease, Nbr, which is responsible for the isoform pattern of miR-34-5p: upon knockdown of nbr, the long isoform accumulates, while the short isoforms decline (Han et al., 2011; Liu et al., 2011). [score:2]
Small RNA Northerns show that the miR-34 pattern is compromised in nbr null. [score:1]
Nibbler is a 3′-to-5′ exonuclease whose function is critical for trimming the 3′end of miR-34-5p in Drosophila (Han et al., 2011; Liu et al., 2011). [score:1]
We first confirmed the effects of Nbr on miRNAs, such as miR-34-5p and miR-305-5p in the ovary libraries. [score:1]
Loss of miR-34 leads to accelerated aging in Drosophila. [score:1]
Some of these defects associated with nbr overlap with those of miR-34-5p (brain degeneration, loss of climbing ability, shorter lifespan); however, whether these phenotypes depend on the loss of 3′end trimming of miRNAs and notably miR-34-5p, or other functions of Nbr on small RNAs, awaits further study. [score:1]
Study of Drosophila miR-34-5p shows that miR-34-5p displays a pattern of different length isoforms due to 3′end heterogeneity and that the short isoform accumulates with age (Liu et al., 2012). [score:1]
Temporally, Nbr showed little change in level with age in the adult head (Fig. 1B), despite the fact that the lower isoform of miR-34-5p accumulates with age (Liu et al., 2012). [score:1]
miR-34-5p trimming was compromised in the nbr null combination, and the Nbr protein level and miR-34-5p trimming pattern were fully rescued by introducing the wild-type genomic nbr transgene (Fig. 2C). [score:1]
However, the miR-34 pattern is rescued by pCaSpeR- nbr (WT) and pCaSpeR- nbr. [score:1]
The DNA oligonucleotides used to make probe templates were miR-34-5p (5′-GAT AAT ACG ACT CAC TAT AGG GAG A-3′/5′-AAA AAA TGG CAG TGT GGT TAG CTG GTT GTG TCT CCC TAT AGT GAG TCG TAT TAT C-3′) and 2S rRNA (5′- GAT AAT ACG ACT CAC TAT AGG GAG A-3′/5′-TGC TTG GAC TAC ATA TGG TTG AGG GTT GTA TCT CCC TAT AGT GAG TCG TAT TAT C-3′). [score:1]
[1 to 20 of 15 sentences]
5
[+] score: 23
For miR-277–3p, miR-987–5p and miR-34–5p their cognate miRNA precursors also increase upon Dis3 knockdown, suggesting that Dis3 normally affects their expression via indirect means, perhaps by increasing the expression of specific transcription factors. [score:7]
[37] Wing phenotypes are not always seen when miRNAs are overexpressed in the wing; for example, no phenotype is seen when miR-34 is overexpressed using MS1096-GAL4. [score:5]
In contrast, the levels of the pri/pre-miRNAs are increased in levels in Dis3 -depleted cells for miR-277–3p, miR-34–5p, miR-317–5p and miR-987–5p suggesting that these miRNAs are not direct targets of Dis3. [score:4]
Using this stringent filtering method, we identified 6 miRNAs whose expression increased ≥2-fold in the knockdown samples compared to both parental controls (miR-277–3p, miR-987–5p, miR-252–5p, miR-34–5p, and miR-7–3p and miR-317–5p). [score:3]
The set of miRNAs that increase ≥2-fold; by RNA-seq upon Dis3 depletion are miR-277–3p, miR-987–5p, miR-252–5p, miR-34–5p and miR-317–5p (Table 2). [score:1]
* miR-317 is located in close proximity to miR-34 and miR-277. [score:1]
The microRNA miR-34 modulates ageing and neurodegeneration in Drosophila. [score:1]
A similar effect is seen for miR-34–5p although the changes in levels are not so pronounced. [score:1]
[1 to 20 of 8 sentences]
6
[+] score: 23
Because the miR-34 muscle phenotype was qualitatively similar to many of the other hits in our flight screen, we wanted to confirm that miR-34 Δ/ Δ and miR-34SP did not cause altered expression of other muscle-expressed miRNAs required for muscle maintenance. [score:5]
Statistical significance was established in this case by comparing the expression values of miR-34 Δ/ Δ to the wild-type control using the NanoStringNorm package in R (t-test, ** P<0.003). [score:3]
These included miR-SPs targeting bantam, miR-1, the K-box family (miR-2b, miR-2c and miR-13b displayed strong phenotypes; miR-2a and miR-13a were flight impaired but fell below our stringent cutoff; ), miR-7, the miR-31 family, miR-34, miR-190, miR-957, miR-986, miR-987 and miR-1001. [score:3]
Our analysis suggests that disruption of miR-34 and 11 other miRNAs can induce a progressive disruption of IFM structure and function, thus uncovering a substantial regulatory landscape for muscle maintenance and/or homeostasis. [score:2]
Thus, we sought to confirm this function for miR-34 by examining a null mutation 42. [score:2]
Indeed, homozygous null animals (miR-34 Δ/ Δ) display age -dependent deficits in flight behaviour that are slightly more severe than miR-34SP at 10 days but reach comparable levels at 30 days (Fig. 4k). [score:1]
Interestingly, loss of Drosophila miR-34 has been reported to induce late-onset brain degeneration 42, raising the intriguing possibility of a general tissue maintenance theme. [score:1]
Recent studies suggest that vertebrate orthologues for several of the conserved miRNAs required for muscle maintenance in our screen (miR-1, miR-7, miR-31 and miR-34, and the K-box orthologue miR-23) are associated with muscle physiology in vertebrate species (Supplementary Table 1). [score:1]
For all other genotypes, statistical significance was established by comparing the miR-SP values, against miR-34 Δ/ Δ, Scramble-SP and wild-type controls. [score:1]
However, to our knowledge, only miR-1 and miR-34 have been implicated by LOF in vertebrate cardiac and/or skeletal muscle function 43 50. [score:1]
Unlike miR-1, miR-34 had not previously been analysed in Drosophila muscle despite mounting evidence implicating this conserved miRNA in muscle function (Supplementary Table 1). [score:1]
Only the levels of mature miR-34–5p were substantially reduced in the null mutant. [score:1]
Thoraces from 1- to 2-day-old adult females of relevant genotypes (dMef2-Gal4>miR34b; dMef2-Gal4>Scramble-SP; miR-34 Δ/ Δ (a gift from Nancy Bonini) and Iso white-1,2,3) were dissected (n≥4) in PBS in biological duplicates. [score:1]
[1 to 20 of 13 sentences]
7
[+] score: 22
Mating causes a reduction in expression of mir-34, mir-92b and mir-988 and an increase in expression of mir-286 (Figure  3; Additional file 4). [score:5]
Females that overexpress mir-34 and mir-92b lay 34% and 37% fewer eggs after mating, whereas females that overexpress mir-286 and mir-988 lay 16% and 33% more eggs than controls (Figure  7). [score:5]
To test the connectivity of the computational networks (Figures  4, 5 and 6) and evaluate causality, we overexpressed six miRNAs for which overexpression lines were available (mir-281-1, mir-286, mir-34, mir-92b, mir-310 and mir-988) using the binary GAL4-UAS expression system [38] under a universal ubiquitin promoter. [score:5]
Number of eggs laid by five young mated females over 18 hours between control and mir-286, mir-34, mir-92b and mir-988 overexpression lines. [score:3]
To assess causality between post-mating modulation of miRNA expression and oviposition, we measured egg laying after mating by control females and females overexpressing mir-286, mir-34, mir-92b, or mir-988. [score:3]
We obtained UAS-mir-281-1, UAS-mir-286, UAS-mir-34, UAS-mir-92b, UAS-mir-988, mir-310 and the co-isogenic control lines from the Bloomington Stock Center. [score:1]
[1 to 20 of 6 sentences]
8
[+] score: 14
Conversely, the significant stimulatory effect of 20E on miR-1-3p and the tendency of 20E to stimulate miR-34-5p that we observed in B. germanica contrasts with the results obtained in D. melanogaster, where 20E did not affect miR-1-3p and inhibited miR-34-5p [30]. [score:3]
miR-14-3p has been described targeting the ecdysone receptor (EcR) in D. melanogaster[29], and miR-34-5p is inducible by the JH analogue methoprene in Drosophila S2 cells [30]. [score:3]
However, the expression levels of miR-1-3p and miR-100-5p were significantly increased after treatment with 20E, whereas those of bantam-3p, miR-125-5p, miR-14-3p, miR-276-3p, miR-34-5p and let-7-5p showed a tendency to increase with respect to controls, although differences were not statistically significant. [score:3]
Two expression bursts of miR-1-3p, miR-34-5p and mir-276-3p occurred in N6, the first just after emergence and the other coinciding or close to the peak of 20E of this stage (Figure  2). [score:3]
Finally, we included two additional miRNAs (miR-14-3p and miR-34-5p) because of their potential interest in the context of moulting and metamorphosis. [score:1]
The differences are also illustrated by the results obtained after JH treatment of B. germanica, which abolished the stimulatory effects of 20E on practically all miRNAs (Figure  3B), which was also the case for miR-34-5p. [score:1]
[1 to 20 of 6 sentences]
9
[+] score: 13
Phylogenetic analysis, target gene prediction and pathway analysis showed that, among the 13 conserved miRNAs (miR-1, miR-100, miR-10a, miR-124, miR-125, miR-184, miR-33, miR-34, miR-7, miR-9, miR-92a, miR-92b and miR-let7), several highly conserved miRNAs (miR-1, miR-7 and miR-34) targeted the same or similar genes leading to the same pathways in shrimp, fruit fly and human (Figure 3b). [score:5]
Some miRNAs, such as miR-34 and miR-S12, could target 7–8 genes. [score:3]
Among the differentially expressed miRNAs found, miR-1, miR-7 and miR-34 are highly conserved and mediate similar pathways, suggesting that some beneficial miRNAs have been preserved in animals during evolution. [score:3]
In our study, phylogenetic analysis showed that the miR-1, miR-7 and miR-34 are highly conserved in shrimp, fruit fly and human and function in similar pathways. [score:1]
Evolutionary analysis showed that three of them, miR-1, miR-7 and miR-34, are highly conserved in shrimp, fruit fly and humans and function in the similar pathways. [score:1]
[1 to 20 of 5 sentences]
10
[+] score: 10
Various combinations of recombinant SmD1 or lysates from SmD1-overexpression cells or SmD1-knockdown cells were subject to microprocessor assay using pri-miR-34 as substrate. [score:3]
Notably, a major SmD1 binding peak directly overlaps with mature miR-34 (Fig 5D), suggesting that SmD1 may directly associate with mature miRNAs (in the context of miRISC). [score:3]
Importantly, levels of several miRNAs, including miR-33, miR-34, miR-276a, miR-317, miR-2b, miR-184 and miR-bantam, were significantly reduced upon SmD1 knockdown (Fig 1B–1G), reminiscent of the phenotype elicited by the loss of the canonical miRNA biogenesis enzyme Drosha. [score:2]
An alternative and non-mutually exclusive possibility is that SmD1 could bind the sequence segment corresponding to mature miR-34 within the primary miRNA transcript. [score:1]
Interestingly, among the primary miRNA transcripts that we have examined so far, miR-33 and miR-34 are the top two highly enriched in immunopurified SmD1 complex (Figs 3C, 5C and 5D). [score:1]
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11
[+] score: 10
Other miRNAs from this paper: dme-mir-275, dme-mir-317
Whereas control flies expressed miR-34 normally, showing three major mature forms, N br [cas9] flies lacked the smaller isoforms with an accumulation of the top isoform, reflecting a trimming defect. [score:3]
Expression of a wild-type Nbr transgene in N br [cas9] flies restored all miR-34 isoforms, indicating a functional rescue. [score:3]
Assessment of miR-34 isoforms confirmed N br [cas9] as a null mutation (Fig.  2D). [score:2]
Interestingly, the levels of miR-34-5p shorter isoforms, which are established Nbr trimming products, were markedly increased in aged ovaries (Fig.  7A). [score:1]
N br [KI-Myc] flies showed a normal miR-34 trimming pattern, suggesting that the addition of a Myc tag has no effect on native protein function (data not shown). [score:1]
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12
[+] score: 9
Although miR-277 is upregulated, miR-34 is downregulated and miR-317 is steadily expressed after HS in the w [1118] strain. [score:9]
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13
[+] score: 7
Other miRNAs from this paper: dme-mir-10, dme-mir-277, dme-mir-190
Further, we found that these target lists overlap (with up to 29 targets) with the well-studied miRNAs mir-34, mir-277, mir-190, and mir-10. [score:5]
Of note, this gene is located in the Drosophila genome in the immediate vicinity (a few hundred basepairs) of mir-34 and mir-277, hinting at a potentially deeper regulatory connection. [score:2]
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14
[+] score: 6
Other miRNAs from this paper: dme-mir-92a, dme-mir-92b, dme-mir-9c, dme-let-7, dme-mir-312
The Eip74EF mRNA is predicted to have three target sites for miR-34 (Table 1; Figure 2A), and miR-34 represses expression of the luc/Eip74EF reporter (Figure 2B). [score:5]
Each primary transcript was amplified by PCR from genomic DNA to generate fragments with the following sequence coordinates: let-7: 2L:18450072-18450291; mir-92b: 3R:21466427-21466673; mir-312: 2R:15647675-15647897; mir-34: 3R:5926642-5926792. [score:1]
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15
[+] score: 6
Both mir-34-5p and let-7-5p were highly abundant in developing embryos, further supporting that they are virtually absent from the unfertilized egg and expressed from the zygotic genome at later stages during development. [score:4]
On the other hand, the conserved microRNA mir-34 has been also described as a maternal microRNA (Soni et al. 2013) but it has only one read copy in our dataset, and it has not been detected in two other independent high-throughput screens (Lee et al. 2014; Ninova et al. 2015). [score:1]
All these findings suggest that either mir-34 is a very low copy maternal microRNA, or that it is rapidly degraded after egg deposition/activation. [score:1]
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16
[+] score: 4
Other miRNAs from this paper: dme-mir-277, dme-bantam, dme-let-7
We verified that in our nibbler knockdowns, miR-34 was stabilized at the length generated by Dicer-1 cleavage (Figure 6A, miR-34 blot, arrowhead). [score:2]
For example, miR-34 is initially produced as a 24 nt product, but Nibbler resects 3–4 nt from its 3′ end. [score:1]
Effective depletion of Dicer-1 was evidenced by the loss of mature miR-144, accumulation of pre-mir-34, and shift in the size distribution of the remaining mature miR-34 to its longest isoform. [score:1]
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17
[+] score: 4
They are let-7, miR-281, miR-34, miR-12, and miR-306. [score:1]
aegypti miR-34 contains a large segment in the loop region, which may be the cause for no score or "NS" by miRscan for the An. [score:1]
aegypti pre-miR-34 alignment. [score:1]
gambiae and D. melanogaster pre-miR-34 alignment was used for miRscan, which produced a score of 17.99.9. [score:1]
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18
[+] score: 3
Other miRNAs from this paper: dme-mir-275, dme-mir-9b, dme-mir-312
Interestingly, the E74 transcription unit displays features common to other potential LARK targets: it contains an A-rich element in the 3′UTR, the transcription unit contains a large ∼37-kb intron, and the 3′UTR contains binding sites for several miRNAs including miR-34, miR-9b, miR312, miR275, and miR-iab-4-5p. [score:3]
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[+] score: 3
Other examples of clustered miRNAs or multicopy miRNAs include: novel miRNA C5152a antisense to C5152b; novel C5303 overlapping ame-mir-137; ame-mir-9b overlapping the ame-mir-79 locus, but on the opposite strand; ame-mir-12 near ame-mir-283; ame-mir-275 near ame-mir-305; ame-mir-277 near ame-mir-317 and ame-mir-34; C1504 near ame-mir-375; and ame-let-7 on the same scaffold as ame-mir-100. [score:1]
Ame-mir-277, ame-mir-317 and ame-mir-34 occur in the same intron of GB10191 - a core component of the RNA polymerase II complex. [score:1]
Ame-mir-34, ame-mir-277 and ame-mir-317 all occupy intron 3 of GB10191. [score:1]
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20
[+] score: 3
They exert the opposite effects on the expression of temporal miRNA genes, such as mir-34, mir-100, mir-125 and let-7 [22]. [score:3]
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21
[+] score: 3
Other miRNAs from this paper: hsa-mir-33a, hsa-mir-92a-1, hsa-mir-92a-2, dme-mir-1, dme-mir-8, dme-mir-11, hsa-mir-34a, hsa-mir-210, dme-mir-184, dme-mir-275, dme-mir-92a, dme-mir-276a, dme-mir-277, dme-mir-33, dme-mir-281-1, dme-mir-281-2, dme-mir-276b, dme-mir-210, dme-mir-92b, dme-bantam, dme-mir-309, dme-mir-317, hsa-mir-1-2, hsa-mir-184, hsa-mir-190a, hsa-mir-1-1, hsa-mir-34b, hsa-mir-34c, aga-bantam, aga-mir-1, aga-mir-184, aga-mir-210, aga-mir-275, aga-mir-276, aga-mir-277, aga-mir-281, aga-mir-317, aga-mir-8, aga-mir-92a, aga-mir-92b, hsa-mir-92b, hsa-mir-33b, hsa-mir-190b, dme-mir-190, dme-mir-957, dme-mir-970, dme-mir-980, dme-mir-981, dme-mir-927, dme-mir-989, dme-mir-252, dme-mir-1000, aga-mir-1174, aga-mir-1175, aga-mir-34, aga-mir-989, aga-mir-11, aga-mir-981, aga-mir-1889, aga-mir-1890, aga-mir-1891, aga-mir-190, aga-mir-927, aga-mir-970, aga-mir-957, aga-mir-1000, aga-mir-309, cqu-mir-1174, cqu-mir-281-1, cqu-mir-1, cqu-mir-275, cqu-mir-957, cqu-mir-277, cqu-mir-252-1, cqu-mir-970, cqu-mir-317-1, cqu-mir-981, cqu-mir-989, cqu-mir-1175, cqu-mir-276-1, cqu-mir-276-2, cqu-mir-276-3, cqu-mir-210, cqu-mir-92, cqu-mir-190-2, cqu-mir-190-1, cqu-mir-1000, cqu-mir-11, cqu-mir-8, cqu-bantam, cqu-mir-1891, cqu-mir-184, cqu-mir-1890, cqu-mir-980, cqu-mir-33, cqu-mir-2951, cqu-mir-2941-1, cqu-mir-2941-2, cqu-mir-2952, cqu-mir-1889, cqu-mir-309, cqu-mir-252-2, cqu-mir-281-2, cqu-mir-317-2, aga-mir-2944a-1, aga-mir-2944a-2, aga-mir-2944b, aga-mir-2945, aga-mir-33, aga-mir-980
gambiae miRNAs, miR-34, miR-1174, miR-1175, and miR-989, show changes in expression during Plasmodium infection [11]. [score:3]
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22
[+] score: 2
To characterize the interaction between the five miRNAs (miR-279-3p, miR-8-3-p, miR-275-3p, miR-34- 3p and miR-304-5p) and the 3′-UTR of bmfrn mRNA, the full length sequence of the 3′-UTR of bmfrn and the sequences of all miRNA candidates were submitted online to RNAhybrid, an algorithm taking into account the free energy level of RNA -RNA duplexes and degree of miRNA target sequence complementarity 21. [score:1]
Mimics of miR-279-3p, miR-8-3-p, miR-275-3p, miR-34-3p and miR-304-5p (sequences of these miRNAs are listed in Table S1) as well as a negative control (NC) were synthesized by GenePharma (Shanghai, China). [score:1]
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23
[+] score: 2
The mir-34 locus is less than 400 bp downstream of dfmr1 and is transcribed in the opposite direction relative to dfmr1. [score:2]
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24
[+] score: 1
The significance of maternal miRNAs pre-MZT is largely unknown, except for dme-miR-34, which is maternally inherited and important for neurogenesis in Drosophila melanogaster [25]. [score:1]
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25
[+] score: 1
The maternal loading of NFE involves conserved miRNAs such as let-7, Bantam, Mir-34, Mir-305, Mir-8, Mir-71 and Mir-1, all of which play roles in basic biological functions [6], as well as large amounts of MIR-bg5 miRNAs. [score:1]
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26
[+] score: 1
Other miRNAs from this paper: dme-bantam, dme-mir-971, dme-mir-986, dme-mir-1012
We observed no editing events in the loop regions, even in those microRNAs that contained significant number of reads (>100) partially (alternative cleavage site) or entirely in loop regions, such as mir-34 (data not shown). [score:1]
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27
[+] score: 1
Similarly, the adult onset of Drosophila miR-34 promotes longevity and maintains neuronal homeostasis by repressing Eip74EF, a transcription factor required for progression through earlier life stages [1, 3]. [score:1]
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28
[+] score: 1
In zebrafish, miR-34 was the only reported maternal miRNA, which was involved in embryonic neurogenesis [32]. [score:1]
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29
[+] score: 1
In this specific instance, miR-2a and miR-34 were identified from Helicoverpa armigera and S. litura based on in silico comparative analysis using data set of B. mori. [score:1]
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30
[+] score: 1
Other miRNAs from this paper: dme-mir-33, dsi-mir-34, dsi-mir-33
The bars show the ratio between the relative number of 21–23 nt small RNAs mapping to each virus (normalised by number of reads of the abundant Drosophila miRNA miR-34-5p), and the number of virus RNA-seq reads (relative to non-viral RNAseq reads, excluding rRNA reads). [score:1]
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31
[+] score: 1
The 277~34 cluster also has intra-cluster TSSs upstream of dme-mir-34. [score:1]
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32
[+] score: 1
There can be two sub-categories delineated; miRNAs that do not change normally under stress, but do in dystrophic mutants (miR-92a and miR-34) and miRNAs that change as a normal response, but do not in Dys and Dg mutants (miR-956, miR-252, miR-970, miR-137, miR-986, miR-193, miR-1017, miR-962, miR-315, miR-1013, miR-980, miR-975, miR-190, miR-iab-4as-5p, miR-1003 and miR-313). [score:1]
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33
[+] score: 1
And ten pre-miRNAs also predicted mature parts on the star (*) arm (mir-305, mir-79, let-7, mir-2a-2, mir-8, mir-7, mir-9a, mir-316, mir-34, mir-12). [score:1]
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34
[+] score: 1
Other miRNAs from this paper: hsa-let-7a-1, hsa-let-7a-2, hsa-let-7a-3, hsa-let-7b, hsa-let-7c, hsa-let-7d, hsa-let-7e, hsa-let-7f-1, hsa-let-7f-2, hsa-mir-27a, hsa-mir-29a, hsa-mir-101-1, dme-mir-1, dme-mir-2a-1, dme-mir-2a-2, dme-mir-2b-1, dme-mir-2b-2, dme-mir-10, mmu-let-7g, mmu-let-7i, mmu-mir-1a-1, mmu-mir-101a, mmu-mir-124-3, mmu-mir-126a, mmu-mir-133a-1, mmu-mir-137, mmu-mir-140, mmu-mir-142a, mmu-mir-155, mmu-mir-10b, mmu-mir-183, mmu-mir-193a, mmu-mir-203, mmu-mir-143, hsa-mir-10a, hsa-mir-10b, hsa-mir-34a, hsa-mir-183, hsa-mir-199b, hsa-mir-203a, hsa-mir-210, hsa-mir-222, hsa-mir-223, dme-mir-133, dme-mir-124, dme-mir-79, dme-mir-210, dme-mir-87, mmu-mir-295, mmu-mir-34c, mmu-mir-34b, mmu-let-7d, dme-let-7, dme-mir-307a, dme-mir-2c, hsa-let-7g, hsa-let-7i, hsa-mir-1-2, hsa-mir-124-1, hsa-mir-124-2, hsa-mir-124-3, hsa-mir-133a-1, hsa-mir-133a-2, hsa-mir-137, hsa-mir-140, hsa-mir-142, hsa-mir-143, hsa-mir-126, hsa-mir-193a, mmu-let-7a-1, mmu-let-7a-2, mmu-let-7b, mmu-let-7c-1, mmu-let-7c-2, mmu-let-7e, mmu-let-7f-1, mmu-let-7f-2, mmu-mir-29a, mmu-mir-27a, mmu-mir-34a, mmu-mir-101b, hsa-mir-1-1, mmu-mir-1a-2, hsa-mir-155, mmu-mir-10a, mmu-mir-210, mmu-mir-223, mmu-mir-222, mmu-mir-199b, mmu-mir-124-1, mmu-mir-124-2, hsa-mir-101-2, hsa-mir-34b, hsa-mir-34c, hsa-mir-378a, mmu-mir-378a, mmu-mir-133a-2, mmu-mir-133b, hsa-mir-133b, mmu-mir-411, hsa-mir-193b, hsa-mir-411, mmu-mir-193b, hsa-mir-944, dme-mir-193, dme-mir-137, dme-mir-994, mmu-mir-1b, mmu-mir-101c, hsa-mir-203b, mmu-mir-133c, mmu-let-7j, mmu-let-7k, mmu-mir-126b, mmu-mir-142b, mmu-mir-124b
Beyond mammals, 20 miRNA families were found conserved across fruitfly, mouse and human, 8 were conserved between fruitfly and worm, and 4 families (let-7, miR-1, miR-34 and miR-124) were found conserved among all the 4 species. [score:1]
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35
[+] score: 1
The level of repression of five low abundance mature microRNAs (let-7-3p, miR-307a-5p, miR-970-5p, miR-1003-5p, miR-34-3p) is not significantly different from the empty control vector (paired t-test; p<0.05). [score:1]
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