Efficient isolation of mouse deletion mutant embryonic stem cells by CRISPR.
STAR Protoc
; 3(2): 101436, 2022 06 17.
Article
en En
| MEDLINE
| ID: mdl-35693210
ABSTRACT
Gene functions can be assessed in mouse embryonic stem (ES) cells and in mutant mice derived from mutant ES cells. Here, we describe an approach for efficient isolation of the ES clones carrying deletion mutations at the target genes by CRISPR-Cas9. Two sgRNAs against a target gene are co-expressed with puromycin-resistant gene in ES cells through co-transfection followed by transient puromycin selection. Deletion mutations are identified by PCR from individual ES clones that are picked from puromycin-selected ES cells.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Sistemas CRISPR-Cas
/
Células Madre Embrionarias de Ratones
Límite:
Animals
Idioma:
En
Revista:
STAR Protoc
Año:
2022
Tipo del documento:
Article
País de afiliación:
China