Developmental Cell
Volume 32, Issue 1, 12 January 2015, Pages 97-108
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Article
Reverse Genetic Screening Reveals Poor Correlation between Morpholino-Induced and Mutant Phenotypes in Zebrafish

https://doi.org/10.1016/j.devcel.2014.11.018Get rights and content
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Highlights

  • Site-specific nucleases used to generate a collection of zebrafish mutants

  • Most mutants displayed normal embryonic development

  • Most mutants failed to recapitulate published Morpholino phenotypes

  • Parallel informatics analysis suggests high false-positive rates for Morpholinos

Summary

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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Co-first author

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Present address: Department of Biomedical Engineering, Khalifa University of Science, Technology and Research (KUSTAR), PO Box 127788, Abu Dhabi, United Arab Emirates

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Present address: Institute of Cell Biology and Neuroscience and Buchmann Institute for Molecular Life Sciences (BMLS), University of Frankfurt, 60438 Frankfurt am Main, Germany