Trio_mouse_sequencing_to_establish_crispr_off_target_rates

This study sequences parent/offspring trios of whole-genome-sequenced C57BL/6 mice, where the parents are wild-type, and the founder embryos are in one of five states: (1) wild-type (untreated) embryos, (2) cytoplasmic sham-injected (MI'd but with Cas9 only) embryos, (3) cytoplasmic sham-injected (MI'd but with water) embryos, (4) mutant (cytoplasmic MI with cas9 + synthetic Tyr grna#1 on Tyr), (5) mutant (cytoplasmic MI with cas9 + synthetic Tyr grna#2 on Tyr). Each offspring will be de-novo variant called, to detect the crispr-induced on-target lesion and the combined effect of any potential off-target mutations and background de-novo mutations. Comparing the off-target + de-novo rate in the crispr-treated embryos to the untreated embryos will allow us to estimate the rate of crispr off-target mutations for the two Tyr- sgRNAs in C56BL/6 mice.