ArticleCell genomics2025
Strategic targeting of Cas9 nickase expands tandem gene arrays.
Article in Cell genomics, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed.
- Technologies for engineering repetitive DNA.Quantitative biology (Beijing, China) · 2026Review
- Parallel gene amplification by Cas9 nickase for generating functionally heterogeneous cell populations.Cell reports methods · 2026Article
- Transcriptional capacity limits copper resistance in yeast harboring highly expandedmicroPublication biology · 2026Article
- Emerging drivers of DNA repeat expansions.Biochemical Society transactions · 2025Review
- Catalytically inactive Cas9 attenuates DNA end resection: A potential application for region-restricted random mutagenesis.iScience · 2025Article
- The Slx4-Rad1-Rad10 nuclease differentially regulates deletions and duplications induced by a replication fork barrier.PLoS genetics · 2025Article
- Long-read plasmid sequencing strategy for evaluating intrinsic instability of tandem gene arrays.microPublication biology · 2025Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
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.
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