Department of Genetics and Biotechnology, St. Petersburg State University, 199034 St. Petersburg, Russia.
Vavilov Institute of General Genetics, St. Petersburg Branch, Russian Academy of Sciences, 199034 St. Petersburg, Russia.
Int J Mol Sci. 2023 Jul 26;24(15):11960. doi: 10.3390/ijms241511960.
The yeast 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.
酵母是一种独特的遗传研究对象，已开发出广泛的相对简单、廉价且耗时短的方法，可用于进行各种基因组修饰。其中包括点突变、特定基因和染色体区域的缺失和破坏、用于表达异源基因的盒插入、靶向染色体重排（如易位和倒位）、有丝分裂类型的定向改变（特定染色体的丢失或复制，倍性水平的改变）、交配型改变等。近年来，CRISPR/Cas9 技术使经典的酵母基因组操作得以推进，该技术可在酵母基因组中同时产生多种变化。在本综述中，我们讨论了经典的酵母基因组修饰方法和 CRISPR/Cas9 技术的实际应用。此外，我们还回顾了倍性变化的方法，包括非整倍体的产生、交配型转换方法和定向 DSB。结合有用的选择标记和转化技术的描述，这项工作几乎代表了酵母基因组修饰的完整指南。
