Effective heritable gene knockdown in zebrafish using synthetic microRNAs

Jean Giacomotto (), Silke Rinkwitz and Thomas S. Becker ()
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Jean Giacomotto: Brain and Mind Research Institute, Sydney Medical School, University of Sydney
Silke Rinkwitz: Brain and Mind Research Institute, Sydney Medical School, University of Sydney
Thomas S. Becker: Brain and Mind Research Institute, Sydney Medical School, University of Sydney

Nature Communications, 2015, vol. 6, issue 1, 1-11

Abstract: Abstract Although zebrafish is used to model human diseases through mutational and morpholino-based knockdown approaches, there are currently no robust transgenic knockdown tools. Here we investigate the knockdown efficiency of three synthetic miRNA-expressing backbones and show that these constructs can downregulate a sensor transgene with different degrees of potency. Using this approach, we reproduce spinal muscular atrophy (SMA) in zebrafish by targeting the smn1 gene. We also generate different transgenic lines, with severity and age of onset correlated to the level of smn1 inhibition, recapitulating for the first time the different forms of SMA in zebrafish. These lines are proof-of-concept that miRNA-based approaches can be used to generate potent heritable gene knockdown in zebrafish.

Date: 2015
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DOI: 10.1038/ncomms8378

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