title: 2C-Cas9: a versatile tool for clonal analysis of gene function.
abstract: CRISPR/Cas9-mediated targeted mutagenesis allows efficient generation of loss-of-function alleles in zebrafish. To date, this technology has been primarily used to generate genetic knockout animals. Nevertheless, the study of the function of certain loci might require tight spatiotemporal control of gene inactivation. Here, we show that tissue-specific gene disruption can be achieved by driving Cas9 expression with the Gal4/UAS system. Furthermore, by combining the Gal4/UAS and Cre/loxP systems, we establish a versatile tool to genetically label mutant cell clones, enabling their phenotypic analysis. Our technique has the potential to be applied to diverse model organisms, enabling tissue-specific loss-of-function and phenotypic characterization of live and fixed tissues.
authors.affiliation: Institut Curie, PSL Research University, INSERM U 934, CNRS UMR3215, F-75005, Paris, France;
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Di Donato
authors.affiliation: Institut Curie, PSL Research University, INSERM U 934, CNRS UMR3215, F-75005, Paris, France;
authors.forename: Flavia
authors.initials: F
authors.lastname: De Santis
authors.affiliation: Institut Curie, PSL Research University, INSERM U 934, CNRS UMR3215, F-75005, Paris, France;
authors.forename: Thomas O
authors.initials: TO
authors.lastname: Auer
authors.affiliation: Institut Curie, PSL Research University, INSERM U 934, CNRS UMR3215, F-75005, Paris, France;
authors.forename: Noé
authors.initials: N
authors.lastname: Testa
authors.affiliation: Department of Cardiovascular Development and Repair, Atherothrombosis and Imaging, Centro Nacional de Investigaciones Cardiovasculares (CNIC), 28028 Madrid, Spain;
authors.forename: Héctor
authors.initials: H
authors.lastname: Sánchez-Iranzo
authors.affiliation: Department of Cardiovascular Development and Repair, Atherothrombosis and Imaging, Centro Nacional de Investigaciones Cardiovasculares (CNIC), 28028 Madrid, Spain;
authors.forename: Nadia
authors.initials: N
authors.lastname: Mercader
authors.affiliation: Muséum National d'Histoire Naturelle, INSERM U1154, CNRS UMR 7196, Paris F-75231, France.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Concordet
authors.affiliation: Institut Curie, PSL Research University, INSERM U 934, CNRS UMR3215, F-75005, Paris, France;
authors.forename: Filippo
authors.identifier: 0000-0001-8551-2846
authors.initials: F
authors.lastname: Del Bene
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D020868:Gene Silencing; D030781:Organisms, Genetically Modified / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism