Engineering APOBEC3A deaminase for highly accurate and efficient base editing.
Nat Chem Biol
; 20(9): 1176-1187, 2024 Sep.
Article
in En
| MEDLINE
| ID: mdl-38553609
ABSTRACT
Cytosine base editors (CBEs) are effective tools for introducing C-to-T base conversions, but their clinical applications are limited by off-target and bystander effects. Through structure-guided engineering of human APOBEC3A (A3A) deaminase, we developed highly accurate A3A-CBE (haA3A-CBE) variants that efficiently generate C-to-T conversion with a narrow editing window and near-background level of DNA and RNA off-target activity, irrespective of methylation status and sequence context. The engineered deaminase domains are compatible with PAM-relaxed SpCas9-NG variant, enabling accurate correction of pathogenic mutations in homopolymeric cytosine sites through flexible positioning of the single-guide RNAs. Dual adeno-associated virus delivery of one haA3A-CBE variant to a mouse model of tyrosinemia induced up to 58.1% editing in liver tissues with minimal bystander editing, which was further reduced through single dose of lipid nanoparticle-based messenger RNA delivery of haA3A-CBEs. These results highlight the tremendous promise of haA3A-CBEs for precise genome editing to treat human diseases.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Cytidine Deaminase
/
Gene Editing
Limits:
Animals
/
Humans
Language:
En
Journal:
Nat Chem Biol
Journal subject:
BIOLOGIA
/
QUIMICA
Year:
2024
Document type:
Article
Affiliation country:
China
Country of publication:
Estados Unidos