ArticlePloS one2017
PCR artifact in testing for homologous recombination in genomic editing in zebrafish.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 34 citations in OpenAlex.
- PCR analysis of insertion sequences leads to the generation of artefact amplicons.Access microbiology · 2026Article
- Efficient non-viral immune cell engineering using circular single-stranded DNA-mediated genomic integration.Nature biotechnology · 2025Article
- Transgene Mapping in Animals: What to Choose?International journal of molecular sciences · 2025Review
- Generation and application of endogenously floxed alleles for cell-specific knockout in zebrafish.Developmental cell · 2023Article
- Efficient single copy integration via homology-directed repair (scHDR) by 5'modification of large DNA donor fragments in mice.Nucleic acids research · 2023Article
- A Critical Review of Zebrafish Neurological Disease Models-1. The Premise: Neuroanatomical, Cellular and Genetic Homology and Experimental Tractability.Oxford open neuroscience · 2023Article
- The Spermidine Synthase GeneCells · 2022Article
- Article
- GeneWeld: Efficient Targeted Integration Directed by Short Homology in Zebrafish.Bio-protocol · 2021Article
- Targeted DNA insertion in plants.Proceedings of the National Academy of Sciences of the United States of America · 2021Review
- CRISPR/Cas9-Mediated Multi-Allelic Gene Targeting in Sugarcane Confers Herbicide Tolerance.Frontiers in genome editing · 2021Article
- A cell cycle-dependent CRISPR-Cas9 activation system based on an anti-CRISPR protein shows improved genome editing accuracy.Communications biology · 2020Article
- Cell Synchronization Enhances Nuclear Transformation and Genome EditingACS synthetic biology · 2020Article
- Production of a mutant of large-scale loach Paramisgurnus dabryanus with skin pigmentation loss by genome editing with CRISPR/Cas9 system.Transgenic research · 2019Article
- CRISPR/Cas9-mediated homology-directed repair by ssODNs in zebrafish induces complex mutational patterns resulting from genomic integration of repair-template fragments.Disease models & mechanisms · 2018Article
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Authors and funding
2 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We report a PCR-induced artifact in testing for homologous recombination in zebrafish. We attempted to replace the lnx2a gene with a donor cassette, mediated by a TALEN induced double stranded cut. The donor construct was flanked with homology arms of about 1 kb at the 5' and 3' ends. Injected embryos (G0) were raised and outcrossed to wild type fish. A fraction of the progeny appeared to have undergone the desired homologous recombination, as tested by PCR using primer pairs extending from genomic DNA outside the homology region to a site within the donor cassette. However, Southern blots revealed that no recombination had taken place. We conclude that recombination happened during PCR in vitro between the donor integrated elsewhere in the genome and the lnx2a locus. We conclude that PCR alone may be insufficient to verify homologous recombination in genome editing experiments in zebrafish.
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