2C-Cas9: a versatile tool for clonal analysis of gene function

• DOI: 10.1101/gr.196170.115
• Publication Date: 2016-03-09T04:08:26Z
• Authors: Vincenzo Di Donato; Flavia De Santis; Thomas O. Auer; Noé Testa; Héctor Sánchez-Ir...; Nadia Mercader; Jean-Paul Concordet; Filippo Del Bene
• Keywords: Gene targeting; Loss function; Gene knockout; Forward genetics; Insertional mutagenesis; Genetic screen

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 function certain loci might require tight spatiotemporal control gene inactivation. Here, we show that tissue-specific disruption can be achieved by driving Cas9 expression with Gal4/UAS system. Furthermore, combining and Cre/loxP systems, establish a versatile tool genetically label mutant cell clones, enabling their phenotypic analysis. Our technique potential applied diverse model organisms, characterization live fixed tissues.