Targeted inhibition of miRNA maturation with morpholinos reveals a role for miR-375 in pancreatic islet development
- PMID: 17676975
- PMCID: PMC1925136
- DOI: 10.1371/journal.pbio.0050203
Targeted inhibition of miRNA maturation with morpholinos reveals a role for miR-375 in pancreatic islet development
Expression of concern in
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Expression of Concern: Targeted Inhibition of miRNA Maturation with Morpholinos Reveals a Role for miR-375 in Pancreatic Islet Development.PLoS Biol. 2022 Apr 29;20(4):e3001631. doi: 10.1371/journal.pbio.3001631. eCollection 2022 Apr. PLoS Biol. 2022. PMID: 35486652 Free PMC article. No abstract available.
Abstract
Several vertebrate microRNAs (miRNAs) have been implicated in cellular processes such as muscle differentiation, synapse function, and insulin secretion. In addition, analysis of Dicer null mutants has shown that miRNAs play a role in tissue morphogenesis. Nonetheless, only a few loss-of-function phenotypes for individual miRNAs have been described to date. Here, we introduce a quick and versatile method to interfere with miRNA function during zebrafish embryonic development. Morpholino oligonucleotides targeting the mature miRNA or the miRNA precursor specifically and temporally knock down miRNAs. Morpholinos can block processing of the primary miRNA (pri-miRNA) or the pre-miRNA, and they can inhibit the activity of the mature miRNA. We used this strategy to knock down 13 miRNAs conserved between zebrafish and mammals. For most miRNAs, this does not result in visible defects, but knockdown of miR-375 causes defects in the morphology of the pancreatic islet. Although the islet is still intact at 24 hours postfertilization, in later stages the islet cells become scattered. This phenotype can be recapitulated by independent control morpholinos targeting other sequences in the miR-375 precursor, excluding off-target effects as cause of the phenotype. The aberrant formation of the endocrine pancreas, caused by miR-375 knockdown, is one of the first loss-of-function phenotypes for an individual miRNA in vertebrate development. The miRNA knockdown strategy presented here will be widely used to unravel miRNA function in zebrafish.
Conflict of interest statement
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References
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- Bernstein E, Kim SY, Carmell MA, Murchison EP, Alcorn H, et al. Dicer is essential for mouse development. Nat Genet. 2003;35:215–217. - PubMed
-
- Giraldez AJ, Cinalli RM, Glasner ME, Enright AJ, Thomson JM, et al. MicroRNAs regulate brain morphogenesis in zebrafish. Science. 2005;308:833–838. - PubMed
-
- Wienholds E, Koudijs MJ, van Eeden FJ, Cuppen E, Plasterk RH. The microRNA-producing enzyme Dicer1 is essential for zebrafish development. Nat Genet. 2003;35:217–218. - PubMed
-
- Ambros V. The functions of animal microRNAs. Nature. 2004;431:350–355. - PubMed
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