ArticleNucleic acids research2020
Enhancing site-specific DNA integration by a Cas9 nuclease fused with a DNA donor-binding domain.
Article in Nucleic acids research, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 35 citations in OpenAlex.
- Gene editing of hematopoietic stem cells: applications and advances.International journal of hematology · 2026Review
- Human Genome Safe Harbor Sites: A Comprehensive Review of Criteria, Discovery, Features, and Applications.Cells · 2026Review
- Transposon-derived genome editors in plants: from compact nucleases to large-fragment integration and regeneration strategies.Engineering in life sciences · 2026Review
- Loss of Alkbh5 enhances AT2 cell differentiation and alveolar repair across diverse injury models via mInternational journal of biological sciences · 2026Article
- Improving Gene Knock-In Efficiencies in Sheep Primary Cells Using a CRISPR/Cas9-Gal4 System.Current issues in molecular biology · 2025Article
- Engineering of BZ transposase and transposon donor vector for enhanced efficiency and safety in gene delivery applications.Nucleic acids research · 2025Article
- exoCasMINI: A T5 exonuclease fused CRISPR-Cas12f system with enhanced gene editing efficiency.iScience · 2025Article
- hpCasMINI: An engineered hypercompact CRISPR-Cas12f system with boosted gene editing activity.Nature communications · 2025Article
- Turning "trashed" genomic loci into treasurable sites for integrating chimeric antigen receptors in T and NK cells.Molecular therapy : the journal of the American Society of Gene Therapy · 2025Review
- Human Pluripotent Stem Cell-Derived Alveolar Organoids for Gene-editing and Lung Adenocarcinomas Modeling.International journal of biological sciences · 2025Article
- Next-generation CRISPR technology for genome, epigenome and mitochondrial editing.Transgenic research · 2024Review
- Precise Gene Knock-In Tools with Minimized Risk of DSBs: A Trend for Gene Manipulation.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Review
- Review
- Engineered circular guide RNAs boost CRISPR/Cas12a- and CRISPR/Cas13d-based DNA and RNA editing.Genome biology · 2023Article
- Phase-separated DropCRISPRa platform for efficient gene activation in mammalian cells and mice.Nucleic acids research · 2023Article
- Enrichment of transgene integrations by transient CRISPR activation of a silent reporter gene.Molecular therapy. Methods & clinical development · 2023Article
- Drag-and-drop genome insertion of large sequences without double-strand DNA cleavage using CRISPR-directed integrases.Nature biotechnology · 2023Article
- Strategies to improve homology-based repair outcomes following CRISPR-based gene editing in mosquitoes: lessons in how to keep any repair disruptions local.Virology journal · 2022Review
- Identification of theGenes · 2022Article
- Transposase-CRISPR mediated targeted integration (TransCRISTI) in the human genome.Scientific reports · 2022Article
Corrections and comments
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Authors and funding
10 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The CRISPR/Cas system is widely used for genome editing. However, robust and targeted insertion of a DNA segment remains a challenge. Here, we present a fusion nuclease (Cas9-N57) to enhance site-specific DNA integration via a fused DNA binding domain of Sleeping Beauty transposase to tether the DNA segment to the Cas9/sgRNA complex. The insertion was unidirectional and specific, and DNA fragments up to 12 kb in length were successfully integrated. As a test of the system, Cas9-N57 mediated the insertion of a CD19-specific chimeric antigen receptor (CD19-CAR) cassette into the AAVS1 locus in human T cells, and induced intrahepatic cholangiocarcinoma in mice by simultaneously mediating the insertion of oncogenic KrasG12D into the Rosa26 locus and disrupting Trp53 and Pten. Moreover, the nuclease-N57 fusion proteins based on AsCpf1 (AsCas12a) and CjCas9 exhibited similar activity. These findings demonstrate that CRISPR-associated nuclease-N57 protein fusion is a powerful tool for targeted DNA insertion and holds great potential for gene therapy applications.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.