Efficient genome editing by CRISPR-Mb3Cas12a in mice.
J Cell Sci
; 133(9)2020 05 11.
Article
em En
| MEDLINE
| ID: mdl-32393674
ABSTRACT
As an alternative and complementary approach to Cas9-based genome editing, Cas12a has not been widely used in mammalian cells largely due to its strict requirement for the TTTV protospacer adjacent motif (PAM) sequence. Here, we report that Mb3Cas12a (Moraxella bovoculi AAX11_00205) can efficiently edit the mouse genome based on the TTV PAM sequence with minimal numbers of large on-target deletions or insertions. When TTTV PAM sequence-targeting CRISPR (cr)RNAs of 23 nt spacers are used, >70% of the founders obtained are edited. Moreover, the use of Mb3Cas12a tagged to monomeric streptavidin (mSA) in conjunction with biotinylated DNA donor template leads to high knock-in efficiency in two-cell mouse embryos, with 40% of founders obtained containing the desired knock-in sequences.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas
/
Edição de Genes
Limite:
Animals
Idioma:
En
Revista:
J Cell Sci
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Estados Unidos