TY - JOUR

T1 - Practical Approaches for the Yeast Saccharomyces cerevisiae Genome Modification

AU - Stepchenkova, Elena I.

AU - Zadorsky, Sergey P.

AU - Shumega, Andrey R.

AU - Aksenova, Anna Y.

PY - 2023/7/26

Y1 - 2023/7/26

N2 - The yeast S. cerevisiae is a unique genetic object for which a wide range of relatively simple, inexpensive, and non-time-consuming methods have been developed that allow the performing of a wide variety of genome modifications. Among the latter, one can mention point mutations, disruptions and deletions of particular genes and regions of chromosomes, insertion of cassettes for the expression of heterologous genes, targeted chromosomal rearrangements such as translocations and inversions, directed changes in the karyotype (loss or duplication of particular chromosomes, changes in the level of ploidy), mating-type changes, etc. Classical yeast genome manipulations have been advanced with CRISPR/Cas9 technology in recent years that allow for the generation of multiple simultaneous changes in the yeast genome. In this review we discuss practical applications of both the classical yeast genome modification methods as well as CRISPR/Cas9 technology. In addition, we review methods for ploidy changes, including aneuploid generation, methods for mating type switching and directed DSB. Combined with a description of useful selective markers and transformation techniques, this work represents a nearly complete guide to yeast genome modification.

AB - The yeast S. cerevisiae is a unique genetic object for which a wide range of relatively simple, inexpensive, and non-time-consuming methods have been developed that allow the performing of a wide variety of genome modifications. Among the latter, one can mention point mutations, disruptions and deletions of particular genes and regions of chromosomes, insertion of cassettes for the expression of heterologous genes, targeted chromosomal rearrangements such as translocations and inversions, directed changes in the karyotype (loss or duplication of particular chromosomes, changes in the level of ploidy), mating-type changes, etc. Classical yeast genome manipulations have been advanced with CRISPR/Cas9 technology in recent years that allow for the generation of multiple simultaneous changes in the yeast genome. In this review we discuss practical applications of both the classical yeast genome modification methods as well as CRISPR/Cas9 technology. In addition, we review methods for ploidy changes, including aneuploid generation, methods for mating type switching and directed DSB. Combined with a description of useful selective markers and transformation techniques, this work represents a nearly complete guide to yeast genome modification.

KW - CRISPR-Cas Systems/genetics

KW - Gene Editing/methods

KW - Saccharomyces cerevisiae/genetics

KW - genome editing

KW - CRISPR/Cas9

KW - genome modifications

KW - yeast transformation

KW - Saccharomyces cerevisiae

KW - yeast

UR - https://www.mendeley.com/catalogue/1793f8d8-ae33-3879-996f-573283be0e08/

U2 - 10.3390/ijms241511960

DO - 10.3390/ijms241511960

M3 - Review article

C2 - 37569333

VL - 24

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

SN - 1422-0067

IS - 15

M1 - 118960

ER -