A multifunctional AAV-CRISPR-Cas9 and its host response.
Nat Methods
; 13(10): 868-74, 2016 10.
Article
in En
| MEDLINE
| ID: mdl-27595405
ABSTRACT
CRISPR-Cas9 delivery by adeno-associated virus (AAV) holds promise for gene therapy but faces critical barriers on account of its potential immunogenicity and limited payload capacity. Here, we demonstrate genome engineering in postnatal mice using AAV-split-Cas9, a multifunctional platform customizable for genome editing, transcriptional regulation, and other previously impracticable applications of AAV-CRISPR-Cas9. We identify crucial parameters that impact efficacy and clinical translation of our platform, including viral biodistribution, editing efficiencies in various organs, antigenicity, immunological reactions, and physiological outcomes. These results reveal that AAV-CRISPR-Cas9 evokes host responses with distinct cellular and molecular signatures, but unlike alternative delivery methods, does not induce extensive cellular damage in vivo. Our study provides a foundation for developing effective genome therapeutics.
Full text:
1
Database:
MEDLINE
Main subject:
Genetic Engineering
/
Gene Transfer Techniques
/
Dependovirus
/
CRISPR-Cas Systems
/
Genetic Vectors
Limits:
Animals
/
Humans
Language:
En
Journal:
Nat Methods
Journal subject:
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Year:
2016
Type:
Article
Affiliation country:
United States