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While CRISPR-Cas9 is a marvelous new addition to our gene editing toolbox, undesired mutations may occur. In mice, off-target editing can be largely managed by choosing high specificity guide RNAs and segregating any off-targets that occur by breeding over several generations. However, mutations at the target locus, such as large deletions, inversions, and chromosomal translocations, can also occur (; ). Recently, donor DNAs, a necessary component for introducing specific sequences into the genome by homology directed repair, have been shown to sometimes integrate in a head-to-tail, or “multiplexed” manner. This type of on-target mutation is often impossible to detect in the F0 generation ( ). For this reason, we now offer an F0 founder breeding and allele sequence validation service that includes testing for the multiplexed integration of donor DNA. We encourage you to use our expertise in validating genome-edited alleles in your next mouse project.
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Released July 13, 2020