Optimized protocol to generate genome-wide inactivated Cas9-expressing murine T cells.
STAR Protoc
; 4(1): 101922, 2023 03 17.
Article
en En
| MEDLINE
| ID: mdl-36516053
ABSTRACT
In vivo genome-wide CRISPR screens in primary T cells allow the systematic and unbiased identification of non-redundant regulatory mechanisms shaping immune responses. Here, we present an optimized protocol for efficient generation of a pool of genome-wide inactivated Cas9-expressing T cells using a retroviral library of sgRNA. We detail the process of large-scale viral production and library integration in activated murine T cells as well as the two-step PCR approach for sgRNA recovery and abundance evaluation. For complete details on the use and execution of this protocol, please refer to Sutra Del Galy et al. (2020).
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Sistemas CRISPR-Cas
/
ARN Guía de Sistemas CRISPR-Cas
Límite:
Animals
Idioma:
En
Revista:
STAR Protoc
Año:
2023
Tipo del documento:
Article
País de afiliación:
Francia