論文 - 詳細
| RRC ID | 63004 |
|---|---|
| 著者 | Yokouchi Y, Suzuki S, Ohtsuki N, Yamamoto K, Noguchi S, Soejima Y, Goto M, Ishioka K, Nakamura I, Suzuki S, Takenoshita S, Era T. |
| タイトル | Rapid repair of human disease-specific single-nucleotide variants by One-SHOT genome editing. |
| ジャーナル | Sci Rep |
| Abstract |
Many human diseases ranging from cancer to hereditary disorders are caused by single-nucleotide mutations in critical genes. Repairing these mutations would significantly improve the quality of life for patients with hereditary diseases. However, current procedures for repairing deleterious single-nucleotide mutations are not straightforward, requiring multiple steps and taking several months to complete. In the current study, we aimed to repair pathogenic allele-specific single-nucleotide mutations using a single round of genome editing. Using high-fidelity, site-specific nuclease AsCas12a/Cpf1, we attempted to repair pathogenic single-nucleotide variants (SNVs) in disease-specific induced pluripotent stem cells. As a result, we achieved repair of the Met918Thr SNV in human oncogene RET with the inclusion of a single-nucleotide marker, followed by absolute markerless, scarless repair of the RET SNV with no detected off-target effects. The markerless method was then confirmed in human type VII collagen-encoding gene COL7A1. Thus, using this One-SHOT method, we successfully reduced the number of genetic manipulations required for genome repair from two consecutive events to one, resulting in allele-specific repair that can be completed within 3 weeks, with or without a single-nucleotide marker. Our findings suggest that One-SHOT can be used to repair other types of mutations, with potential beyond human medicine. |
| 巻・号 | 10(1) |
| ページ | 13927 |
| 公開日 | 2020-8-18 |
| DOI | 10.1038/s41598-020-70401-7 |
| PII | 10.1038/s41598-020-70401-7 |
| PMID | 32811847 |
| PMC | PMC7435196 |
| MeSH | Alleles Bacterial Proteins / genetics Bacterial Proteins / metabolism* CRISPR-Associated Proteins / genetics CRISPR-Associated Proteins / metabolism* CRISPR-Cas Systems / genetics Collagen Type VII / genetics Collagen Type VII / metabolism Endodeoxyribonucleases / genetics Endodeoxyribonucleases / metabolism* Endonucleases / genetics Gene Editing / methods* Genome, Human / genetics Humans Induced Pluripotent Stem Cells / physiology Mutation / genetics Nucleotides / genetics Pluripotent Stem Cells / physiology Polymorphism, Single Nucleotide / genetics* Proto-Oncogene Proteins c-ret / genetics Proto-Oncogene Proteins c-ret / metabolism |
| IF | 3.998 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 10 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | 409B2(HPS0076) |