ArticlePlant biotechnology journal2025
Cas9-Rep fusion tethers donor DNA in vivo and boosts the efficiency of HDR-mediated genome editing.
Article in Plant biotechnology journal, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- Writing Big in Plant Genomes: Advances, Challenges and Strategies for Targeted Large-Fragment DNA Insertion.Plant, cell & environment · 2026Review
- Transposase-Assisted Donor Tethering Boosts Large-Fragment HDR in Plants.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- From gametocides to gene drive: natural transmission distortion systems as tools for cereal breeding.Journal of applied genetics · 2026Review
- CRISPR/Cas Genome Editing and Its Applications in Cereal Crop Improvement.Plant-environment interactions (Hoboken, N.J.) · 2026Review
- Multilayer regulation of CRISPR systems: integrating anti-CRISPR proteins, CRISPRi/a, and quorum sensing networks.Frontiers in cellular and infection microbiology · 2026Review
- The evolving landscape of precise DNA insertion in plants.Nature communications · 2025Review
- Advances and prospects of large DNA fragment editing in plants.Nature plants · 2025Review
- Recent advances in site-specific transgene insertion into the maize genome using recombinases and genome editing endonucleases.Frontiers in plant science · 2025Review
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
Genome editing based on the homology-directed repair (HDR) pathway enables scar-free and precise genetic manipulations. However, the low frequency of HDR hinders its application in plant genome editing. In this study, we engineered the fusion of Cas9 and a viral replication protein (Rep) as a molecular bridge to tether donor DNA in vivo, which enhances the efficiency of targeted gene insertion via the HDR pathway. This Rep-bridged knock-in (RBKI) method combines the advantages of rolling cycle replication of viral replicons and in vivo enrichment of donor DNA at the target site for HDR. Chromatin immunoprecipitation indicated that the Cas9-Rep fusion protein bound up to 66-fold more donor DNA than Cas9 did. We exemplified the RBKI method by inserting small- to middle-sized tags (33-519 bp) into 3 rice genes. Compared to Cas9, Cas9-Rep fusion increased the KI frequencies by 4-7.6-fold, and up to 72.2% of stable rice transformants carried in-frame knock-in events in the T
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Registered trials
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