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2 publications mentioning dre-mir-722

Open access articles that are associated with the species Danio rerio and mention the gene name mir-722. Click the [+] symbols to view sentences that include the gene name, or the word cloud on the right for a summary.

1
[+] score: 220
Consistent with our data that miR-722 directly downregulates endogenous rac2 expression, defects in neutrophil motility (Fig.  3B) or their recruitment to tissue injury (Fig.  3C) or infection (Fig.  3D; Movie 3) resulting from miR-722 overexpression were all rescued by rac2 overexpression, pinpointing rac2 as a relevant miR-722 target in neutrophils. [score:13]
miR-722 directly suppresses zebrafish r ac2 expressionWe then confirmed that miR-722 can directly suppress the zebrafish r ac2 gene. [score:9]
Furthermore, the motility of the miR-722 -overexpressing neutrophils was significantly hampered (Fig.  1F; Movie 2), which phenocopied the Rac2 -deficient neutrophils (Deng et al., 2011; Rosowski et al., 2016), coinciding with the prediction that miR-722 downregulates rac2 expression in neutrophils. [score:8]
To further validate that rac2 is a major target of miR-722 in neutrophils, we performed a rescue experiment using a transgenic zebrafish line that overexpresses zebrafish rac2 followed by the SV40 3′UTR, which is resistant to miR-722 -mediated suppression (Deng et al., 2011). [score:7]
Restoring rac2 expression in the miR-722 -overexpressing line increased susceptibility, to levels comparable to the wild-type larvae, from the acute systemic Pseudomonas infection (Fig.  4G), suggesting that miR-722 protects zebrafish from lethal inflammatory challenge via the suppression of rac2. [score:7]
Furthermore, miR-722 -overexpressing larvae display improved outcomes in both sterile and bacterial systemic mo dels, which correlates with a robust upregulation of the anti-inflammatory cytokines in the whole larvae and isolated neutrophils. [score:6]
miR-722 directly suppresses zebrafish r ac2 expression. [score:6]
The miR-722 -overexpressing larvae showed increased survival compared with those overexpressing the vector control (Fig.  4A), despite the presence of similar bacterial burdens (possibly as a result of intact macrophage functions), excluding the possibility that the miR-722 -overexpressing line had increased bactericidal activity (Fig.  4B). [score:6]
We observed specific upregulation of both the precursor and mature forms of miR-722 in the transgenic animals, without alterations in the level of miR-223 or a ubiquitously expressed miRNA, let-7e (Fig.  1B), confirming that miRNA biogenesis in neutrophils is intact. [score:6]
Fig. 3. Overexpression of rac2 rescues neutrophil recruitment in miR-722 -expressing larvae. [score:5]
Expression of miR-722 significantly suppressed the relative luciferase activity, which was dependent on the seed sequences in the zebrafish r ac2 gene (Fig.  2B). [score:5]
We performed reporter assays to validate the direct translational suppression by miR-722. [score:5]
To demonstrate that the miR-722 mimic also elicits its protective role via the inhibition of rac2 in neutrophils, the mimic was injected into a line that expressed miR-722-resistant rac2 in neutrophils. [score:5]
Indeed, the protective role of miR-722 was abrogated in the rac2 -overexpressing line, but not in a line that expresses the mCherry control in neutrophils (Fig.  6D). [score:5]
Similar expression changes of inos2b, il-10 and tgf-β2 were observed in FACS-isolated neutrophils, consistent with neutrophil-restricted overexpression of miR-722 (Fig.  4F). [score:5]
In the miR-722 -overexpressing zebrafish line, the rac2 mRNA level is significantly reduced (Fig.  2C), suggesting a direct destabilization of the rac2 transcript by miR-722 in neutrophils. [score:4]
We then confirmed that miR-722 can directly suppress the zebrafish r ac2 gene. [score:4]
miR-722 downregulates the transcript level of rac2 through binding to seed-matching sequence in the rac2 3′UTR. [score:4]
In wild-type larvae, the endogenous level of miR-722 was not upregulated during systemic inflammation (Fig.  4H). [score:4]
Fig. 2. miR-722 downregulates the zebrafish r ac2 transcript through binding to seed complementary sequences in its 3′UTR. [score:4]
In addition, despite the fact that anti-inflammatory cytokines were upregulated in infected larvae that received miR-722 mimic, the fold changes were variable between experiments that did not reach statistical significance (Fig.  6F,G). [score:4]
Fig. 5. The miR-722 -overexpressing line has increased resistance to sterile systemic inflammation. [score:3]
Here, we have identified a miRNA, miR-722, that, when overexpressed in neutrophils, reduces neutrophil chemotaxis and protects the whole organism from both sterile and non-sterile inflammatory assaults. [score:3]
First, we generated a transgenic zebrafish line that overexpresses miR-722 specifically in neutrophils (schematic in Fig.  1A). [score:3]
To facilitate the identification and characterization of cells expressing this miRNA, a 206 bp genomic DNA sequence flanking miRNA-722 was cloned into an intron that allows co -expression of miR-722 with a green fluorescent reporter protein, Dendra2. [score:3]
Thus, we tested whether miR-722 -overexpressing zebrafish were more resistant to lethal inflammatory challenges; in this experiment, we used a bacterial systemic infection mo del using the Gram -negative bacteria Pseudomonas aeruginosa PAK strain. [score:3]
In comparison, miR-722 -overexpressing larvae survived significantly better (Fig.  5A). [score:3]
All mimics and the miR-722 inhibitor were synthesized by Thermo Fisher Scientific. [score:3]
Rac2 overexpression rescues miR-722 -induced phenotypes. [score:3]
Significantly fewer neutrophils were recruited in miR-722 -overexpressing lines in both incidences (Fig.  1D,E; Movie 1). [score:3]
We performed bioinformatics analysis (TargetScanFish) and identified three miRNAs (miR-194, miR-722 and miR-129) that are predicted to bind to the 3′UTR of both transcript variants of the zebrafish r ac2 gene. [score:3]
Three founders each of the zebrafish that express the vector control or miR-722 were obtained. [score:3]
In contrast, miR-722 overexpression in neutrophils protected zebrafish from lethal challenge of Pseudomonas infection. [score:3]
The pro-inflammatory cytokines, including TNF-α, IL-6 and IL-8, were also induced in the miR-722 -overexpressing line, but not significantly. [score:3]
Embryos at the one-cell stage were injected with 1 nl of 15 µM dre-miR-722 mimic (#4464066), dre-miR-722 inhibitor (#4464084), hsa-miR-129-5p mimic (#4464084), 1 µM dre-miR-223 mimic (#4464066) or buffer as a control. [score:3]
Fig. 1. Neutrophil recruitment and motility is hindered in the miR-722 -overexpressing zebrafish line. [score:3]
Neutrophil-specific miR-722 overexpression protects zebrafish from lethal systemic inflammation. [score:3]
Fig. 4. The miR-722 -overexpressing line has increased resistance to bacterial -induced systemic inflammation. [score:3]
Embryos were injected with buffer, miRNA mimic or an miR-722 inhibitor at the one-cell stage and experiments were performed with larvae at 2 dpf. [score:3]
We have generated a transgenic zebrafish line that overexpresses microRNA-722 (miR-722) in neutrophils. [score:3]
Because zebrafish nos2b also harbors miR-722 -binding sites, we next examined whether rac2 overexpression mitigates the protective effect elicited by miR-722. [score:3]
We next examined the recruitment of miR-722 -overexpressing neutrophils in two separate acute inflammation mo dels: a localized bacterial infection and tail transection. [score:3]
Here, we have demonstrated that rac2 is a major target of miR-722 in neutrophils. [score:3]
miR-722 -overexpressing neutrophils are defective in motility and chemotaxis. [score:3]
As expected, neutrophil recruitment to the injury site was impaired in the larvae receiving miR-722 mimic as compared to the buffer -injected larvae, whereas recruitment was more robust in miR-722 -inhibitor -injected larvae (Fig.  6A). [score:2]
miR-722 and miR-129 share the same seed sequence and bind to a perfect seed-matching site in the rac2 3′UTR with a context+ score percentile above 90. miR-194 binds to a separate site with a partial seed match and a context+ score percentile of 69, possibly being a weaker regulator of rac2. [score:2]
We observed significantly increased resistance to lethal LPS challenge in the miR-722 -mimic -injected larvae compared with buffer- or the miR-722 -inhibitor -injected larvae (Fig.  6B), suggesting that miR-722 expression is a potential prophylactic measure in sterile inflammation. [score:2]
miR-722 expression vector was cloned by amplifying the 722 hairpin from the lyzC:miR-722 vector used to create the transgenic line and inserted into pcDNA3.1 at the HindIII/ XbaI cloning sites using the following primers: pcDNA-722+: 5′- GTTTAAACTTAAGCTTGCCACCATGGATGAGGAAATCGC-3′; pcDNA-722-: 5′-AAACGGGCCCTCTAGAGACCGGTACCCCCGGGCTGC-3′. [score:2]
Tg(lyzC:Dendra2) [pu7] was generated using the same configuration without the miR-722 insertion (vector). [score:1]
Plasmids were deposited to Addgene (plasmid 97158, pSi-check2-zRac2 3′UTR; plasmid 97159, pSi-check2-zRac2 - mut 3′UTR; plasmid 97160, pSi-check2-hRac2 3′UTR; plasmid 97161, pSi-check2-hRac2 - mut 3′UTR; plasmid 97163, miR-722-Dendra pCDNA). [score:1]
By contrast, a protective effect from miR-722 mimic in the Pseudomonas sepsis mo del was not observed (Fig.  6E). [score:1]
For control, Tg(lyzC:miR-722/Dendra2) [pu6] was crossed with Tg(mpx:mCherry) (mCherry). [score:1]
miR-722 mimic protects against sterile inflammation. [score:1]
More than three founders (F0) for both Tg(lyzC:miR-722/Dendra2) [pu6] and Tg(LyzC:Dendra2) [pu7] were obtained as described in the AB background (Deng et al., 2011). [score:1]
The zebrafish rac2 3′UTR harbors a miR-722 -binding site with perfect seed-sequence match (Fig.  2A). [score:1]
Data compiled from previous miRNA sequencing experiments suggest that miR-722 is intergenic and predominantly produces a mature 3p strand that harbors the rac2 -binding sequence (www. [score:1]
Another neutrophil-specific gene, encoding lysozyme C, was not altered in the same sample, indicating the specificity of miR-722 towards rac2. [score:1]
Fig. 6. miR-722 mimic reduces neutrophilic inflammation and mortality from systemic LPS challenge. [score:1]
F2 larvae from Tg(lyzC:miR-722/Dendra2) [pu6] (miR-722) and Tg(lyzC:miR-722/Dendra2) [pu6] (vector) at 3 dpf were injected intravenously with 1000 CFU of Pseudomonas. [score:1]
Finally, we injected an miR-722 mimic into zebrafish embryos at the one-cell stage to deliver miR-722 ubiquitously. [score:1]
Owing to current technical hurdles that prevent us from effectively delivering miR-772 into neutrophils in the larvae, we have not been able to show the efficacy of miR-722 in treating existing inflammation. [score:1]
The plasmids were deposited to Addgene (plasmid 97101, Tol2-lyzC-Vector-Dendra2; plasmid 97130, Tol2-lyzC-miR-722-Dendra2). [score:1]
miR-129-5p, which is conserved between zebrafish and human, shares the same seed sequence as miR-722. [score:1]
Finally, an miR-722 mimic protects zebrafish from lethal lipopolysaccharide challenge. [score:1]
The different host outcome with miR-722 -mimic treatment between sterile and bacterial infection is possibly due to a threshold concentration lower than that of biological relevance or the short-lived effectiveness of the miR-722 mimic to have a sustained effect when battling live organisms that take days to clear (Fig.  4B). [score:1]
In addition, the miR-722 level is below the detection limit by quantitative miRNA reverse-transcription (RT)-PCR (RT-qPCR) in sorted neutrophils. [score:1]
FACS of dissociated embryo neutrophils and one-stepLarvae at 3 dpf from Tg(lyzC:miR-722/Dendra2) [pu6] and Tg(LyzC:Dendra2) [pu7] were injected with 1000 CFU P. aeruginosa (PAK) into the tail vein and incubated to 8 hours post-infection (hpi). [score:1]
Larvae at 3 dpf from Tg(lyzC:miR-722/Dendra2) [pu6] and Tg(LyzC:Dendra2) [pu7] were injected with 1000 CFU P. aeruginosa (PAK) into the tail vein and incubated to 8 hours post-infection (hpi). [score:1]
In addition, similar numbers of neutrophils were present in both lines (Fig.  1C), indicating that miR-722 does not impair neutrophil biogenesis or survival. [score:1]
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[+] score: 1
Other miRNAs from this paper: hsa-let-7a-1, hsa-let-7a-2, hsa-let-7a-3, hsa-let-7b, hsa-let-7e, hsa-mir-20a, hsa-mir-21, hsa-mir-23a, hsa-mir-24-1, hsa-mir-24-2, hsa-mir-26b, hsa-mir-27a, hsa-mir-29a, hsa-mir-31, hsa-mir-29b-1, hsa-mir-29b-2, hsa-mir-103a-2, hsa-mir-103a-1, hsa-mir-199a-1, hsa-mir-148a, hsa-mir-7-1, hsa-mir-7-2, hsa-mir-7-3, hsa-mir-10b, hsa-mir-181a-2, hsa-mir-181b-1, hsa-mir-181c, hsa-mir-199a-2, hsa-mir-199b, hsa-mir-203a, hsa-mir-204, hsa-mir-212, hsa-mir-181a-1, hsa-mir-221, hsa-mir-23b, hsa-mir-27b, hsa-mir-128-1, hsa-mir-132, hsa-mir-133a-1, hsa-mir-133a-2, hsa-mir-143, hsa-mir-200c, hsa-mir-181b-2, hsa-mir-128-2, hsa-mir-200a, hsa-mir-30e, hsa-mir-148b, hsa-mir-338, hsa-mir-133b, dre-mir-7b, dre-mir-7a-1, dre-mir-7a-2, dre-mir-10b-1, dre-mir-181b-1, dre-mir-181b-2, dre-mir-199-1, dre-mir-199-2, dre-mir-199-3, dre-mir-203a, dre-mir-204-1, dre-mir-181a-1, dre-mir-221, dre-mir-222a, dre-let-7a-1, dre-let-7a-2, dre-let-7a-3, dre-let-7a-4, dre-let-7a-5, dre-let-7a-6, dre-let-7b, dre-let-7e, dre-mir-7a-3, dre-mir-10b-2, dre-mir-20a, dre-mir-21-1, dre-mir-21-2, dre-mir-23a-1, dre-mir-23a-2, dre-mir-23a-3, dre-mir-23b, dre-mir-24-4, dre-mir-24-2, dre-mir-24-3, dre-mir-24-1, dre-mir-26b, dre-mir-27a, dre-mir-27b, dre-mir-29b-1, dre-mir-29b-2, dre-mir-29a, dre-mir-30e-2, dre-mir-101b, dre-mir-103, dre-mir-128-1, dre-mir-128-2, dre-mir-132-1, dre-mir-132-2, dre-mir-133a-2, dre-mir-133a-1, dre-mir-133b, dre-mir-133c, dre-mir-143, dre-mir-148, dre-mir-181c, dre-mir-200a, dre-mir-200c, dre-mir-203b, dre-mir-204-2, dre-mir-338-1, dre-mir-338-2, dre-mir-454b, hsa-mir-181d, dre-mir-212, dre-mir-181a-2, hsa-mir-551a, hsa-mir-551b, dre-mir-31, dre-mir-724, dre-mir-725, dre-mir-735, dre-mir-740, hsa-mir-103b-1, hsa-mir-103b-2, dre-mir-2184, hsa-mir-203b, dre-mir-7146, dre-mir-181a-4, dre-mir-181a-3, dre-mir-181a-5, dre-mir-181b-3, dre-mir-181d, dre-mir-204-3, dre-mir-24b, dre-mir-7133, dre-mir-128-3, dre-mir-7132, dre-mir-338-3
Four miRNAs in miRBase, miR-722, miR-735 and miR-740 and miR-7146, were excluded from the analysis as their mapped genomic locations conflicted with miRBase annotation. [score:1]
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