bioRxivGeneticsPreprint
CRISPR-LRS for mapping transgenes in the mouse genome
William Bart BryantiD, Allison C. Yang, Susan H. GriffiniD, Wei ZhangiD, Xiaochun LongiD, Joseph M. MianoiD
Abstract
Microinjected transgenes, including bacterial artificial chromosomes (BACs), insert randomly in the mouse genome. Traditional methods of mapping a transgene are challenging, thus complicating breeding strategies and the accurate interpretation of phenotypes, particularly when a transgene disrupts critical coding or noncoding sequences. Here, we introduce CRISPR-Cas9 long-read sequencing (CRISPR-LRS) to ascertain transgene integration locus and estimated copy number. This method revealed integration loci for both a BAC and Cre-driver line, and estimated the copy numbers for two other BAC mouse lines. CRISPR-LRS offers an easy approach to establish robust breeding practices and accurate phenotyping of most any transgenic mouse line.
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