Generation of dual-gRNA library for combinatorial CRISPR screening of synthetic lethal gene pairs.
STAR Protoc
; 3(3): 101556, 2022 09 16.
Article
en En
| MEDLINE
| ID: mdl-36060092
ABSTRACT
Combinatorial CRISPR screening is useful for investigating synthetic lethality (SL) gene pairs. Here, we detail the steps for dual-gRNA library construction, with the introduction of two backbones, LentiGuide_DKO and LentiCRISPR_DKO. We describe steps for in vitro screening with 22Rv1-Cas9 and SaOS2-Cas9 cells followed by sequencing and data analysis. By introducing two backbones, we optimized the library construction process, facilitated standard pair-end sequencing, and provided options of screening on cells with or without modification of Cas9 expression.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
ARN Guía de Kinetoplastida
/
Sistemas CRISPR-Cas
Tipo de estudio:
Diagnostic_studies
/
Screening_studies
Idioma:
En
Año:
2022
Tipo del documento:
Article