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

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.

journal_name

Genome Res

journal_title

Genome research

authors

Di Donato V,De Santis F,Auer TO,Testa N,Sánchez-Iranzo H,Mercader N,Concordet JP,Del Bene F

doi

10.1101/gr.196170.115

subject

Has Abstract

pub_date

2016-05-01 00:00:00

pages

681-92

issue

5

eissn

1088-9051

issn

1549-5469

pii

gr.196170.115

journal_volume

26

pub_type

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