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Effective heritable gene knockdown in zebrafish using synthetic microRNAs

Author

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  • Jean Giacomotto

    (Brain and Mind Research Institute, Sydney Medical School, University of Sydney
    Present address: Institute of Molecular Bioscience, The University of Queensland, St Lucia, Queensland 4072, Australia)

  • Silke Rinkwitz

    (Brain and Mind Research Institute, Sydney Medical School, University of Sydney
    Sydney Medical School, University of Sydney)

  • Thomas S. Becker

    (Brain and Mind Research Institute, Sydney Medical School, University of Sydney
    Sydney Medical School, University of Sydney)

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.

Suggested Citation

  • Jean Giacomotto & Silke Rinkwitz & Thomas S. Becker, 2015. "Effective heritable gene knockdown in zebrafish using synthetic microRNAs," Nature Communications, Nature, vol. 6(1), pages 1-11, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8378
    DOI: 10.1038/ncomms8378
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