Protocol for precision editing of endogenous Chlamydomonas reinhardtii genes with CRISPR-Cas.
STAR Protoc
; 5(1): 102774, 2024 Mar 15.
Article
en En
| MEDLINE
| ID: mdl-38096061
ABSTRACT
CRISPR-Cas genome engineering in the unicellular green algal model Chlamydomonas reinhardtii has until recently suffered from low integration efficiencies despite traditional genetics being well established. Here, we present a protocol for efficient homology-directed knockin mutagenesis in all commonly used strains of Chlamydomonas. We describe steps for scarless integration of fusion tags and sequence modifications of almost all proteins without the need for a preceding mutant line. We further empower this genetic-editing approach by efficient crossing and highly robust screening protocols. For complete details on the use and execution of this protocol, please refer to Nievergelt et al. (2023).1.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Chlamydomonas reinhardtii
/
Sistemas CRISPR-Cas
Idioma:
En
Revista:
STAR Protoc
Año:
2024
Tipo del documento:
Article