Format

Send to

Choose Destination
Genome Res. 2016 May;26(5):681-92. doi: 10.1101/gr.196170.115. Epub 2016 Mar 8.

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

Author information

1
Institut Curie, PSL Research University, INSERM U 934, CNRS UMR3215, F-75005, Paris, France;
2
Department of Cardiovascular Development and Repair, Atherothrombosis and Imaging, Centro Nacional de Investigaciones Cardiovasculares (CNIC), 28028 Madrid, Spain;
3
Muséum National d'Histoire Naturelle, INSERM U1154, CNRS UMR 7196, Paris F-75231, France.

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.

PMID:
26957310
PMCID:
PMC4864464
DOI:
10.1101/gr.196170.115
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center