論文 - 詳細
| RRC ID | 81881 |
|---|---|
| 著者 | Morisaka H, Yoshimi K, Okuzaki Y, Gee P, Kunihiro Y, Sonpho E, Xu H, Sasakawa N, Naito Y, Nakada S, Yamamoto T, Sano S, Hotta A, Takeda J, Mashimo T. |
| タイトル | CRISPR-Cas3 induces broad and unidirectional genome editing in human cells. |
| ジャーナル | Nat Commun |
| Abstract |
Although single-component Class 2 CRISPR systems, such as type II Cas9 or type V Cas12a (Cpf1), are widely used for genome editing in eukaryotic cells, the application of multi-component Class 1 CRISPR has been less developed. Here we demonstrate that type I-E CRISPR mediates distinct DNA cleavage activity in human cells. Notably, Cas3, which possesses helicase and nuclease activity, predominantly triggered several thousand base pair deletions upstream of the 5'-ARG protospacer adjacent motif (PAM), without prominent off-target activity. This Cas3-mediated directional and broad DNA degradation can be used to introduce functional gene knockouts and knock-ins. As an example of potential therapeutic applications, we show Cas3-mediated exon-skipping of the Duchenne muscular dystrophy (DMD) gene in patient-induced pluripotent stem cells (iPSCs). These findings broaden our understanding of the Class 1 CRISPR system, which may serve as a unique genome editing tool in eukaryotic cells distinct from the Class 2 CRISPR system. |
| 巻・号 | 10(1) |
| ページ | 5302 |
| 公開日 | 2019-12-6 |
| DOI | 10.1038/s41467-019-13226-x |
| PII | 10.1038/s41467-019-13226-x |
| PMID | 31811138 |
| PMC | PMC6897959 |
| MeSH | CRISPR-Associated Proteins / classification CRISPR-Associated Proteins / genetics* CRISPR-Associated Proteins / metabolism CRISPR-Cas Systems / genetics* Clustered Regularly Interspaced Short Palindromic Repeats DNA Cleavage DNA Helicases / metabolism Exons Gene Editing / methods* Gene Expression Regulation / genetics Gene Knockout Techniques / methods HEK293 Cells Humans Induced Pluripotent Stem Cells Muscular Dystrophy, Duchenne / genetics Sequence Deletion |
| IF | 12.121 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 63 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 6.0 |
| リソース情報 | |
| 遺伝子材料 | pCAG-All-in-one-hCascade (RDB18212) pPB-CAG-hCas3 (RDB18213) pBS-U6-crRNA-empty (RDB18214) |