title: Effective heritable gene knockdown in zebrafish using synthetic microRNAs.
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.
authors.affiliation: Brain and Mind Research Institute, Sydney Medical School, University of Sydney, Camperdown, New South Wales 2050, Australia.
authors.forename: Jean
authors.initials: J
authors.lastname: Giacomotto
authors.affiliation: Brain and Mind Research Institute, Sydney Medical School, University of Sydney, Camperdown, New South Wales 2050, Australia.|Department of Physiology, Sydney Medical School, University of Sydney, Camperdown, New South Wales 2050, Australia.
authors.forename: Silke
authors.initials: S
authors.lastname: Rinkwitz
authors.affiliation: Brain and Mind Research Institute, Sydney Medical School, University of Sydney, Camperdown, New South Wales 2050, Australia.|Department of Physiology, Sydney Medical School, University of Sydney, Camperdown, New South Wales 2050, Australia.
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Becker
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D030801:Animals, Genetically Modified; D055785:Gene Knockdown Techniques; D035683:MicroRNAs / Q000235:genetics*; D015027:Zebrafish / Q000235:genetics*; D029961:Zebrafish Proteins / Q000235:genetics
chemical_list: D020413:3' Untranslated Regions; D035683:MicroRNAs; D029961:Zebrafish Proteins