Overcoming the Limitations of CRISPR-Cas9 Systems in <i>Saccharomyces cerevisiae</i>: Off-Target Effects, Epigenome, and Mitochondrial Editing
Modification of the genome of the yeast <i>Saccharomyces cerevisiae</i> 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 a...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-04-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/11/4/1040 |
| _version_ | 1797604141649887232 |
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| author | Genki Sato Kouichi Kuroda |
| author_facet | Genki Sato Kouichi Kuroda |
| author_sort | Genki Sato |
| collection | DOAJ |
| description | Modification of the genome of the yeast <i>Saccharomyces cerevisiae</i> 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. |
| first_indexed | 2024-03-11T04:42:10Z |
| format | Article |
| id | doaj.art-0aaf44a3fe984881b778ab9ee4ff3319 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-11T04:42:10Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-0aaf44a3fe984881b778ab9ee4ff33192023-11-17T20:34:10ZengMDPI AGMicroorganisms2076-26072023-04-01114104010.3390/microorganisms11041040Overcoming the Limitations of CRISPR-Cas9 Systems in <i>Saccharomyces cerevisiae</i>: Off-Target Effects, Epigenome, and Mitochondrial EditingGenki Sato0Kouichi Kuroda1Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, JapanDivision of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, JapanModification of the genome of the yeast <i>Saccharomyces cerevisiae</i> 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.https://www.mdpi.com/2076-2607/11/4/1040<i>Saccharomyces cerevisiae</i>genome editingCRISPR-Cas9 systemCRISPR Nickaseepigeneticsmitochondrial DNA |
| spellingShingle | Genki Sato Kouichi Kuroda Overcoming the Limitations of CRISPR-Cas9 Systems in <i>Saccharomyces cerevisiae</i>: Off-Target Effects, Epigenome, and Mitochondrial Editing Microorganisms <i>Saccharomyces cerevisiae</i> genome editing CRISPR-Cas9 system CRISPR Nickase epigenetics mitochondrial DNA |
| title | Overcoming the Limitations of CRISPR-Cas9 Systems in <i>Saccharomyces cerevisiae</i>: Off-Target Effects, Epigenome, and Mitochondrial Editing |
| title_full | Overcoming the Limitations of CRISPR-Cas9 Systems in <i>Saccharomyces cerevisiae</i>: Off-Target Effects, Epigenome, and Mitochondrial Editing |
| title_fullStr | Overcoming the Limitations of CRISPR-Cas9 Systems in <i>Saccharomyces cerevisiae</i>: Off-Target Effects, Epigenome, and Mitochondrial Editing |
| title_full_unstemmed | Overcoming the Limitations of CRISPR-Cas9 Systems in <i>Saccharomyces cerevisiae</i>: Off-Target Effects, Epigenome, and Mitochondrial Editing |
| title_short | Overcoming the Limitations of CRISPR-Cas9 Systems in <i>Saccharomyces cerevisiae</i>: Off-Target Effects, Epigenome, and Mitochondrial Editing |
| title_sort | overcoming the limitations of crispr cas9 systems in i saccharomyces cerevisiae i off target effects epigenome and mitochondrial editing |
| topic | <i>Saccharomyces cerevisiae</i> genome editing CRISPR-Cas9 system CRISPR Nickase epigenetics mitochondrial DNA |
| url | https://www.mdpi.com/2076-2607/11/4/1040 |
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