Fig. 5.

Expedited biallelic gene editing by FACS sorting. a Schematic of the FACS sorting protocol. GFP^neg / mCh^pos cells (targeted) are isolated in bulk and subjected to nuclease transfection, followed by sorting populations for mCh^neg (excised). Resolved alleles were screened in the population, or in single-cell derived iPSC clones. The donor vector, allele and additional features are as described in Fig. 4a,b. b Representative FACS plot for the targeting and excision steps. GFP^pos cells were excluded, and mCh^pos cells were divided into high and low fractions to bias monoallelically and biallelically targeted cells, respectively. c Predicted APRT allele spectrum within the sorted mCh^neg populations. d APRT diploid genotypes of clones. Only clones with Normal or MMEJ-resolved alleles are shown. Alleles marked as ‘APRT*J’ were also edited with the Silent mutation. e By employing imperfect μH, MhAX derives edited cells and their concordant isogenic controls simultaneously, reducing technical variation