Reverse genetic screening reveals poor correlation between morpholino-induced and mutant phenotypes in zebrafish.

Publisher: Cell Press Country of Publication: United States NLM ID: 101120028 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1878-1551 (Electronic) Linking ISSN: 15345807 NLM ISO Abbreviation: Dev Cell Subsets: MEDLINE

Original Publication: Cambridge, Mass. : Cell Press, c2001-

Deoxyribonucleases/*genetics ; Gene Expression Regulation/*drug effects ; Gene Knockdown Techniques/*methods ; Morpholinos/*pharmacology ; Mutation/*genetics ; Oligonucleotides, Antisense/*pharmacology ; Zebrafish/*genetics ; Zebrafish Proteins/*genetics; Animals ; Blotting, Western ; Deoxyribonucleases/metabolism ; Gene Expression Regulation, Developmental ; Phenotype ; RNA, Messenger/genetics ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Zebrafish/growth & development ; Zebrafish Proteins/antagonists & inhibitors

The widespread availability of programmable site-specific nucleases now enables targeted gene disruption in the zebrafish. In this study, we applied site-specific nucleases to generate zebrafish lines bearing individual mutations in more than 20 genes. We found that mutations in only a small proportion of genes caused defects in embryogenesis. Moreover, mutants for ten different genes failed to recapitulate published Morpholino-induced phenotypes (morphants). The absence of phenotypes in mutant embryos was not likely due to maternal effects or failure to eliminate gene function. Consistently, a comparison of published morphant defects with the Sanger Zebrafish Mutation Project revealed that approximately 80% of morphant phenotypes were not observed in mutant embryos, similar to our mutant collection. Based on these results, we suggest that mutant phenotypes become the standard metric to define gene function in zebrafish, after which Morpholinos that recapitulate respective phenotypes could be reliably applied for ancillary analyses.
(Copyright © 2015 Elsevier Inc. All rights reserved.)

Comment in: Zebrafish. 2015 Oct;12(5):319. (PMID: 26436467)
Comment in: Nat Rev Mol Cell Biol. 2019 Apr;20(4):197. (PMID: 30733603)

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R01 HL093766 United States HL NHLBI NIH HHS; R01HL093766 United States HL NHLBI NIH HHS; R01HL079266 United States HL NHLBI NIH HHS; T32 HD007505 United States HD NICHD NIH HHS; R01 HL101374 United States HL NHLBI NIH HHS; T32 HL120823 United States HL NHLBI NIH HHS; R01HL101374 United States HL NHLBI NIH HHS; R01 HL079266 United States HL NHLBI NIH HHS

0 (Morpholinos)
0 (Oligonucleotides, Antisense)
0 (RNA, Messenger)
0 (Zebrafish Proteins)
EC 3.1.- (Deoxyribonucleases)

Date Created: 20141224 Date Completed: 20150310 Latest Revision: 20220309

20221213

PMC4487878

10.1016/j.devcel.2014.11.018

25533206