Whole genome sequencing for GOTI analysis

A mixture of Cre, Cas9 mRNA, genome targeting sgRNA, CasRX mRNA and RNA targeting sgRNA (targeting Polq as experimental group and scramble sgRNA as control group) into one blastomere of a 2-cell mouse embryo, derived from mTmG male mice mating with wild-type female mice. The action of Cre, injected into one of two cells, is expected to generate chimeric embryo labeled with both GFP and RFP. When the chimeric embryo reaches embryonic day (E14.5), it is minced into small pieces and digested into single-cell suspension. We collected GFP+ and RFP+ cells by fluorescence-activated cell sorting (FACS), respectively. Next, the two population cells are independently processed for whole genome sequence (WGS), which are subsequently processed by standard pipeline.