Directed evolution of CRISPR-Cas9 to increase its specificity.
Nat Commun
; 9(1): 3048, 2018 08 06.
Article
in En
| MEDLINE
| ID: mdl-30082838
ABSTRACT
The use of CRISPR-Cas9 as a therapeutic reagent is hampered by its off-target effects. Although rationally designed S. pyogenes Cas9 (SpCas9) variants that display higher specificities than the wild-type SpCas9 protein are available, these attenuated Cas9 variants are often poorly efficient in human cells. Here, we develop a directed evolution approach in E. coli to obtain Sniper-Cas9, which shows high specificities without killing on-target activities in human cells. Unlike other engineered Cas9 variants, Sniper-Cas9 shows WT-level on-target activities with extended or truncated sgRNAs with further reduced off-target activities and works well in a preassembled ribonucleoprotein (RNP) format to allow DNA-free genome editing.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Directed Molecular Evolution
/
CRISPR-Cas Systems
Limits:
Humans
Language:
En
Journal:
Nat Commun
Journal subject:
BIOLOGIA
/
CIENCIA
Year:
2018
Document type:
Article