Marker-free genetic manipulations in yeast using CRISPR/CAS9 system.
Curr Genet
; 64(5): 1129-1139, 2018 Oct.
Article
em En
| MEDLINE
| ID: mdl-29626221
ABSTRACT
The budding yeast is currently one of the major model organisms for the study of a wide variety of biological processes. Genetic manipulation of yeast involves the extensive usage of selectable markers that can lead to undesired effects. Thus, marker-free genetic manipulation in yeast is highly desirable for gene/promoter replacement and various other applications. Here we combine the power of selectable markers followed by CRISPR/CAS9 genome editing for common genetic manipulations in yeast in a marker-free manner. We demonstrate our approach for whole gene and promoter replacements and for high-efficiency operator array integration. Our approach allows the utilization of many thousands of existing strains including library strains for the generation of significant genetic changes in yeast in a marker-free and cloning-free fashion.
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MEDLINE
Assunto principal:
Saccharomyces cerevisiae
/
Sistemas CRISPR-Cas
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article