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Dissection of zebrafish shha function using site-specific targeting with a Cre-dependent genetic switch

eLife, ISSN: 2050-084X, Vol: 6
2017
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Article Description

Despite the extensive use of zebrafish as a model organism in developmental biology and regeneration research, genetic techniques enabling conditional analysis of gene function are limited. In this study, we generated Zwitch, a Cre-dependent invertible gene-trap cassette, enabling the establishment of conditional alleles in zebrafish by generating intronic insertions via in vivo homologous recombination. To demonstrate the utility of Zwitch, we generated a conditional sonic hedgehog a (shha) allele. Homozygous shha mutants developed normally; however, shha mutant embryos globally expressing Cre exhibited strong reductions in endogenous shha and shha target gene mRNA levels and developmental defects associated with null shha mutations. Analyzing a conditional shha mutant generated using an epicardium-specific inducible Cre driver revealed unique roles for epicardium-derived Shha in myocardial proliferation during heart development and regeneration. Zwitch will extend the utility of zebrafish in organ development and regeneration research and might be applicable to other model organisms.

Bibliographic Details

Sugimoto, Kotaro; Hui, Subhra P; Sheng, Delicia Z; Kikuchi, Kazu

eLife Sciences Organisation, Ltd.

Neuroscience; Biochemistry, Genetics and Molecular Biology; Immunology and Microbiology

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