Efficient shRNA-Mediated Inhibition of Gene Expression in Zebrafish
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Sive_Efficient shRNA.pdf
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Author(s)
Sive, Hazel L.
Rienzo, Gianluca De
Gutzman, Jennifer H.
Date Issued
July 2012
Journal
Zebrafish
Publisher
Mary Ann Liebert
Citation
De Rienzo, Gianluca, Jennifer H. Gutzman, and Hazel Sive. “Efficient shRNA-Mediated Inhibition of Gene Expression in Zebrafish.” Zebrafish 9.3 (2012): 97–107. ©2012 Mary Ann Liebert, Inc. publishers.
Version
Final published version
Abstract
Despite the broad repertoire of loss of function (LOF) tools available for use in the zebrafish, there remains a need for a simple and rapid method that can inhibit expression of genes at later stages. RNAi would fulfill that role, and a previous report (Dong et al. 2009) provided encouraging data. The goal of this study was to further address the ability of expressed shRNAs to inhibit gene expression. This included quantifying RNA knockdown, testing specificity of shRNA effects, and determining whether tissue-specific LOF could be achieved. Using an F0 transgenic approach, this report demonstrates that for two genes, wnt5b and zDisc1, each with described mutant and morphant phenotypes, shRNAs efficiently decrease endogenous RNA levels. Phenotypes elicited by shRNA resemble those of mutants and morphants, and are reversed by expression of cognate RNA, further demonstrating specificity. Tissue-specific expression of zDisc1 shRNAs in F0 transgenics demonstrates that conditional LOF can be readily obtained. These results suggest that shRNA expression presents a viable approach for rapid inhibition of zebrafish gene expression.
MIT Department
Massachusetts Institute of Technology. School of Science
Whitehead Institute for Biomedical Research
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1089/zeb.2012.0770
