2C-Cas9: a versatile tool for clonal analysis of gene function
- Vincenzo Di Donato1,
- Flavia De Santis1,
- Thomas Auer1,
- Noe Testa1,
- Hector Sánchez-Iranzo2,
- Nadia Mercader2,
- Jean-Paul Concordet3 and
- Filippo Del Bene1,4
- 1 Institut Curie;
- 2 Centro Nacional de Investigaciones Cardiovasculares;
- 3 Muséum National d'Histoire naturelle
- ↵* Corresponding author; email: filippo.del-bene{at}curie.fr
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.
- Received June 22, 2015.
- Accepted February 24, 2016.
- Published by Cold Spring Harbor Laboratory Press
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