ArticleNucleic acids research2020
DNA barcoding reveals that injected transgenes are predominantly processed by homologous recombination in mouse zygote.
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 18 papers.
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Who cites it
18 citing papers in PubMed, 35 citations in OpenAlex.
- Unbiased Long-Read Whole-Genome Sequencing Enables High-Resolution Mapping of Transgene Concatenation and Off-target Genomic Disruption in a Mouse Model.Computational and structural biotechnology journal · 2026Article
- CRISPR-Cas9 HDR optimization: RAD52, denatured, and 5'-modified DNA templates in knock-in mice generation.iScience · 2025Article
- Transgene Mapping in Animals: What to Choose?International journal of molecular sciences · 2025Review
- Efficient single copy integration via homology-directed repair (scHDR) by 5'modification of large DNA donor fragments in mice.Nucleic acids research · 2023Article
- Comparative Pre-Clinical Analysis of CD20-Specific CAR T Cells Encompassing 1F5-, Leu16-, and 2F2-Based Antigen-Recognition Moieties.International journal of molecular sciences · 2023Article
- A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair.Scientific reports · 2022Article
- Evaluation of the α-casein (CSN1S1) locus as a potential target for a site-specific transgene integration.Scientific reports · 2022Article
- Efficient targeted transgenesis of large donor DNA into multiple mouse genetic backgrounds using bacteriophage Bxb1 integrase.Scientific reports · 2022Article
- Characterization of Mammalian In Vivo Enhancers Using Mouse Transgenesis and CRISPR Genome Editing.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Review
- Induction of a chromatin boundary in vivo upon insertion of a TAD border.PLoS genetics · 2021Article
- Pannexin 1 Transgenic Mice: Human Diseases and Sleep-Wake Function Revision.International journal of molecular sciences · 2021Article
- Novel transgenic mice with Cre-dependent co-expression of GFP and human ACE2: a safe tool for study of COVID-19 pathogenesis.Transgenic research · 2021Article
- A Comprehensive Review of Genetically Engineered Mouse Models for Prader-Willi Syndrome Research.International journal of molecular sciences · 2021Review
- DGK and DZHK position paper on genome editing: basic science applications and future perspective.Basic research in cardiology · 2021Review
- Interaction of ncRNA and Epigenetic Modifications in Gastric Cancer: Focus on Histone Modification.Frontiers in oncology · 2021Review
- A homology independent sequence replacement strategy in human cells using a CRISPR nuclease.Open biology · 2021Article
- On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes.International journal of molecular sciences · 2020Article
Corrections and comments
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Authors and funding
8 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Mechanisms that ensure repair of double-strand DNA breaks (DSBs) are instrumental in the integration of foreign DNA into the genome of transgenic organisms. After pronuclear microinjection, exogenous DNA is usually found as a concatemer comprising multiple co-integrated transgene copies. Here, we investigated the contribution of various DSB repair pathways to the concatemer formation. We injected mouse zygotes with a pool of linear DNA molecules carrying unique barcodes at both ends and obtained 10 transgenic embryos with 1-300 transgene copies. Sequencing the barcodes allowed us to assign relative positions to the copies in concatemers and detect recombination events that occurred during integration. Cumulative analysis of approximately 1,000 integrated copies reveals that over 80% of them underwent recombination when their linear ends were processed by synthesis-dependent strand annealing (SDSA) or double-strand break repair (DSBR). We also observed evidence of double Holliday junction (dHJ) formation and crossing over during the concatemer formations. Sequencing indels at the junctions between copies shows that at least 10% of DNA molecules introduced into the zygotes are ligated by non-homologous end joining (NHEJ). Our barcoding approach, verified with Pacific Biosciences Single Molecule Real-Time (SMRT) long-range sequencing, documents high activity of homologous recombination after DNA microinjection.
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