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    Microinjected transgenes, both large and small, are known to insert randomly into the mouse genome. Traditional methods of mapping a transgene are challenging, thus complicating breeding strategies and accurate interpretation of phenotypes, particularly when a transgene disrupts critical coding or noncoding sequences. As the vast majority of transgenic mouse lines remain unmapped, we developed CRISPR-Cas9 Long-Read Sequencing (CRISPR-LRS) to ascertain transgene integration loci. This novel approach mapped a wide size range of transgenes and uncovered more complex transgene-induced host genome re-arrangements than previously appreciated. CRISPR-LRS offers a facile, informative approach to establish robust breeding practices and will enable researchers to study a gene without confounding genetic issues. Finally, CRISPR-LRS will find utility in rapidly and accurately interrogating gene/genome editing fidelity in experimental and clinical settings.

    Citation

    W Bart Bryant, Allison Yang, Susan H Griffin, Wei Zhang, Ashiq M Rafiq, Weiping Han, Ferenc Deak, Mary Katherine Mills, Xiaochun Long, Joseph M Miano. CRISPR-Cas9 Long-Read Sequencing for Mapping Transgenes in the Mouse Genome. The CRISPR journal. 2023 Apr;6(2):163-175

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    PMID: 37071672

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