Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants.
Science
; 368(6488): 290-296, 2020 04 17.
Article
en En
| MEDLINE
| ID: mdl-32217751
ABSTRACT
Manipulation of DNA by CRISPR-Cas enzymes requires the recognition of a protospacer-adjacent motif (PAM), limiting target site recognition to a subset of sequences. To remove this constraint, we engineered variants of Streptococcus pyogenes Cas9 (SpCas9) to eliminate the NGG PAM requirement. We developed a variant named SpG that is capable of targeting an expanded set of NGN PAMs, and we further optimized this enzyme to develop a near-PAMless SpCas9 variant named SpRY (NRN and to a lesser extent NYN PAMs). SpRY nuclease and base-editor variants can target almost all PAMs, exhibiting robust activities on a wide range of sites with NRN PAMs in human cells and lower but substantial activity on those with NYN PAMs. Using SpG and SpRY, we generated previously inaccessible disease-relevant genetic variants, supporting the utility of high-resolution targeting across genome editing applications.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Marcación de Gen
/
Predisposición Genética a la Enfermedad
/
Sistemas CRISPR-Cas
/
Edición Génica
/
Proteína 9 Asociada a CRISPR
Límite:
Humans
Idioma:
En
Revista:
Science
Año:
2020
Tipo del documento:
Article
País de afiliación:
Estados Unidos