Overcoming the Limitations of CRISPR-Cas9 Systems in Saccharomyces cerevisiae: Off-Target Effects, Epigenome, and Mitochondrial Editing

Modification of the genome of the yeast Saccharomyces cerevisiae has great potential for application in biological research and biotechnological advancements, and the CRISPR-Cas9 system has been increasingly employed for these purposes. The CRISPR-Cas9 system enables the precise and simultaneous modification of any genomic region of the yeast to a desired sequence by altering only a 20-nucleotide sequence within the guide RNA expression constructs. However, the conventional CRISPR-Cas9 system has several limitations. In this review, we describe the methods that were developed to overcome these limitations using yeast cells. We focus on three types of developments: reducing the frequency of unintended editing to both non-target and target sequences in the genome, inducing desired changes in the epigenetic state of the target region, and challenging the expansion of the CRISPR-Cas9 system to edit genomes within intracellular organelles such as mitochondria. These developments using yeast cells to overcome the limitations of the CRISPR-Cas9 system are a key factor driving the advancement of the field of genome editing.

Automated summary

Modification of the yeast Saccharomyces cerevisiae genome using the CRISPR-Cas9 system has great potential in biological research and biotechnology. Overcoming the limitations of the conventional system, this review focuses on developments in reducing unintended editing and inducing desired changes in the target region's epigenetic state. Moreover, the expansion of the CRISPR-Cas9 system to edit genomes within intracellular organelles like mitochondria is discussed, highlighting the key role of yeast cells in driving advancements in genome editing.