論文 - 詳細
| RRC ID | 85187 |
|---|---|
| 著者 | Takesue H, Okada S, Doi G, Sugiyama Y, Kusumoto E, Ito T. |
| タイトル | Strategic targeting of Cas9 nickase expands tandem gene arrays. |
| ジャーナル | Cell Genom |
| Abstract |
Expanding tandem gene arrays facilitates adaptation through dosage effects and gene family formation via sequence diversification. However, experimental induction of such expansions remains challenging. Here, we introduce a method termed break-induced replication (BIR)-mediated tandem repeat expansion (BITREx) to address this challenge. BITREx places Cas9 nickase adjacent to a tandem gene array to break the replication fork that has just replicated the array, forming a single-ended double-strand break. This break is subsequently end-resected to become single stranded. Since there is no repeat unit downstream of the break, the single-stranded DNA often invades an upstream unit to initiate ectopic BIR, resulting in array expansion. BITREx has successfully expanded gene arrays in budding yeast, with the CUP1 array reaching ∼1 Mb. Furthermore, appropriate splint DNAs allow BITREx to generate tandem arrays de novo from single-copy genes. We have also demonstrated BITREx in mammalian cells. Therefore, BITREx will find various unique applications in genome engineering. |
| 巻・号 | 5(4) |
| ページ | 100811 |
| 公開日 | 2025-4-9 |
| DOI | 10.1016/j.xgen.2025.100811 |
| PII | S2666-979X(25)00067-9 |
| PMID | 40118067 |
| PMC | PMC12008805 |
| MeSH | Animals CRISPR-Associated Protein 9* / metabolism CRISPR-Cas Systems* / genetics DNA Replication / genetics Deoxyribonuclease I* / genetics Deoxyribonuclease I* / metabolism Humans Saccharomyces cerevisiae / genetics Tandem Repeat Sequences* / genetics |
| 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 | 13 |
| 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 | 293T(RCB2202) |