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bioRxiv · 10.1101/2022.01.05.475144

CRISPR-LRS for mapping transgenes in the mouse genome

Bryant, W. B.·Yang, A.·Griffin, S.·Zhang, W.·Long, X.·Miano, J. M.
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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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Bryant, W. B., Yang, A., Griffin, S., Zhang, W., Long, X., Miano, J. M.. 2022-01-06. CRISPR-LRS for mapping transgenes in the mouse genome. https://doi.org/10.1101/2022.01.05.475144

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