Reverse genetic screening reveals poor correlation between Morpholino-induced and mutant phenotypes in zebrafish
By Fatma Kok, M. Shin, C. Ni, Ankit Gupta, Ann S. Grosse, A. V. Impel, B. Kirchmaier, J. Peterson-Maduro, George Kourkoulis, I. Male and 7 more. Developmental Cell, 2014
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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 twenty 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 eighty percent 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.
Subfields: Cell Biology, Molecular Biology
View Paper: https://www.semanticscholar.org/paper/cef653744b271c75736bee5e126f0a52ea948642
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Citations in the corpus
- Zebrafish: Development of a Vertebrate Model Organism (2018, background): “…produced by morpholinos were unable to be replicated by other gene editing techniques, perhaps due to significant off-target effects of morpholinos or different mechanisms of compensation between antisense-mediated knockdown and loss-of-function mutations (Kok et al., 2015; Rossi et al., 2015).”
- Loss-of-function approaches in comparative physiology: is there a future for knockdown experiments in the era of genome editing? (2019, background): “…Somatic cell Juvenile Quadalti et al., 2018 In vitro cell/tissue culture Rainbow trout (Oncorhynchus mykiss) RNAi Lipofection of cultured gill cells – – Kolosov and Kelly, 2017; Kolosov et al., 2017 Laboratory rat (Rattus norvegicus) RNAi Reversible chemical permeabilization of cultured brain…”