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

• Published in: Microorganisms (Basel) Vol. 11; no. 4; p. 1040
• Main Authors: Sato, Genki, Kuroda, Kouichi
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 16.04.2023; MDPI
• Subjects: Biological research; CRISPR; CRISPR Nickase; CRISPR-Cas9 system; Editing; Epigenetics; Gene expression; Gene sequencing; genome editing; Genomes; Mitochondria; Mitochondrial DNA; Mutation; Nucleotide sequence; Nucleotides; Organelles; Review; Saccharomyces cerevisiae; Yeast; Yeasts; genome editing; CRISPR Nickase; mitochondrial DNA; Saccharomyces cerevisiae; CRISPR-Cas9 system; epigenetics; DNA/RNA hybrid
• ISSN: 2076-2607; 2076-2607
• DOI: 10.3390/microorganisms11041040

Modification of the genome of the yeast 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.