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Dev Cell. 2015 Jan 12;32(1):97-108. doi: 10.1016/j.devcel.2014.11.018. Epub 2014 Dec 18.

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

Author information

1
Program in Gene Function and Expression, University of Massachusetts Medical School, Worcester, MA 01605, USA.
2
Hubrecht Institute, 3584CT Utrecht, the Netherlands.
3
Department of Immunology, Genetics, and Pathology, Rudbeck Laboratory, Uppsala University, 751 85 Uppsala, Sweden; Department of Medicinal Biochemistry and Biophysics, Karolinska Institute, 171 77 Stockholm, Sweden.
4
Hubrecht Institute, 3584CT Utrecht, the Netherlands; Institute for Cardiovascular Organogenesis and Regeneration, Cells-in-Motion Cluster of Excellence, University of Münster, 48149 Münster, Germany.
5
Program in Gene Function and Expression, University of Massachusetts Medical School, Worcester, MA 01605, USA; Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
6
Program in Gene Function and Expression, University of Massachusetts Medical School, Worcester, MA 01605, USA. Electronic address: nathan.lawson@umassmed.edu.

Abstract

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.

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PMID:
25533206
PMCID:
PMC4487878
DOI:
10.1016/j.devcel.2014.11.018
[Indexed for MEDLINE]
Free PMC Article
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