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

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...

Full description

Saved in:
Bibliographic Details
Published inGenome research Vol. 26; no. 5; pp. 681 - 692
Main Authors Di Donato, Vincenzo, De Santis, Flavia, Auer, Thomas O, Testa, Noé, Sánchez-Iranzo, Héctor, Mercader, Nadia, Concordet, Jean-Paul, Del Bene, Filippo
Format Journal Article
LanguageEnglish
Published United States Cold Spring Harbor Laboratory Press 01.05.2016
Subjects
Animals
CRISPR-Cas Systems
Danio rerio
Gene Silencing
Method
Organisms, Genetically Modified - genetics
Organisms, Genetically Modified - metabolism
Zebrafish - genetics
Zebrafish - metabolism
Online AccessGet full text

Cover

More Information
Summary: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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
These authors contributed equally to this work.
Present address: Center for Integrative Genomics, Faculty of Biology and Medicine, University of Lausanne, 1015 Lausanne, Switzerland
ISSN:1088-9051
1549-5469
DOI:10.1101/gr.196170.115