CRISPR/Cas9 mediates efficient conditional mutagenesis in Drosophila.
G3 (Bethesda)
; 4(11): 2167-73, 2014 Sep 05.
Article
in En
| MEDLINE
| ID: mdl-25193494
ABSTRACT
Existing transgenic RNA interference (RNAi) methods greatly facilitate functional genome studies via controlled silencing of targeted mRNA in Drosophila. Although the RNAi approach is extremely powerful, concerns still linger about its low efficiency. Here, we developed a CRISPR/Cas9-mediated conditional mutagenesis system by combining tissue-specific expression of Cas9 driven by the Gal4/upstream activating site system with various ubiquitously expressed guide RNA transgenes to effectively inactivate gene expression in a temporally and spatially controlled manner. Furthermore, by including multiple guide RNAs in a transgenic vector to target a single gene, we achieved a high degree of gene mutagenesis in specific tissues. The CRISPR/Cas9-mediated conditional mutagenesis system provides a simple and effective tool for gene function analysis, and complements the existing RNAi approach.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Mutagenesis
/
Gene Targeting
/
Drosophila
/
CRISPR-Cas Systems
Limits:
Animals
Language:
En
Journal:
G3 (Bethesda)
Year:
2014
Document type:
Article
Affiliation country:
China
Publication country:
ENGLAND
/
ESCOCIA
/
GB
/
GREAT BRITAIN
/
INGLATERRA
/
REINO UNIDO
/
SCOTLAND
/
UK
/
UNITED KINGDOM