Reference - Detail
| RRC ID | 85187 |
|---|---|
| Author | Takesue H, Okada S, Doi G, Sugiyama Y, Kusumoto E, Ito T. |
| Title | Strategic targeting of Cas9 nickase expands tandem gene arrays. |
| Journal | 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. |
| Volume | 5(4) |
| Pages | 100811 |
| Published | 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 |
| Altmetric score |
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 13 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | 293T(RCB2202) |