CRISPR-Cas9 Long-Read Sequencing for Mapping Transgenes in the Mouse Genome.

ABSTRACT: 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 gen

SUBMITTER: Bryant WB 

PROVIDER: S-EPMC10123806 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature