Highly efficient targeted chromosome deletions using CRISPR/Cas9.
Biotechnol Bioeng
; 112(5): 1060-4, 2015 May.
Article
in En
| MEDLINE
| ID: mdl-25362885
ABSTRACT
The CRISPR/Cas9 system has emerged as an intriguing new technology for genome engineering. It utilizes the bacterial endonuclease Cas9 which, when delivered to eukaryotic cells in conjunction with a user-specified small guide RNA (gRNA), cleaves the chromosomal DNA at the target site. Here we show that concurrent delivery of gRNAs designed to target two different sites in a human chromosome introduce DNA double-strand breaks in the chromosome and give rise to targeted deletions of the intervening genomic segment. Predetermined genomic DNA segments ranging from several-hundred base pairs to 1 Mbp can be precisely deleted at frequencies of 1-10%, with no apparent correlation between the size of the deleted fragment and the deletion frequency. The high efficiency of this technique holds promise for large genomic deletions that could be useful in generation of cell and animal models with engineered chromosomes.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
DNA
/
Chromosome Deletion
/
RNA, Guide, Kinetoplastida
/
Gene Targeting
/
DNA Breaks, Double-Stranded
/
Clustered Regularly Interspaced Short Palindromic Repeats
Limits:
Humans
Language:
En
Journal:
Biotechnol Bioeng
Year:
2015
Type:
Article
Affiliation country:
United kingdom