ZFIN ID: ZDB-PUB-160310-6
2C-Cas9: a versatile tool for clonal analysis of gene function
Di Donato, V., De Santis, F., Auer, T., Testa, N., Sánchez-Iranzo, H., Mercader, N., Concordet, J.P., Del Bene, F.
Date: 2016
Source: Genome research   26(5): 681-92 (Journal)
Registered Authors: Auer, Thomas, Del Bene, Filippo, De Santis, Flavia, Mercader Huber, Nadia, Testa, Noé
Keywords: none
MeSH Terms:
  • Animals
  • CRISPR-Cas Systems*
  • Gene Silencing*
  • Organisms, Genetically Modified*/genetics
  • Organisms, Genetically Modified*/metabolism
  • Zebrafish*/genetics
  • Zebrafish*/metabolism
PubMed: 26957310 Full text @ Genome Res.
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.