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

Di Donato, Vincenzo; De Santis, Flavia; Auer, Thomas; Testa, Noe; Sánchez-Iranzo, Hector; Mercader Huber, Nadia; Concordet, Jean-Paul; Del Bene, Filippo (2016). 2C-Cas9: a versatile tool for clonal analysis of gene function. Genome research, 26(5), pp. 681-692. Cold Spring Harbor, N.Y 10.1101/gr.196170.115

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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.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Anatomy

UniBE Contributor:

Mercader Huber, Nadia

Subjects:

600 Technology > 610 Medicine & health
500 Science > 570 Life sciences; biology

ISSN:

1549-5469

Publisher:

Cold Spring Harbor, N.Y

Language:

English

Submitter:

Nadia Isabel Mercader Huber

Date Deposited:

30 Jun 2016 16:40

Last Modified:

09 Sep 2016 02:30

Publisher DOI:

10.1101/gr.196170.115

PubMed ID:

26957310

BORIS DOI:

10.7892/boris.79611

URI:

https://boris.unibe.ch/id/eprint/79611

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