Fig. 2

UDiTaS in practice. a. Editing a population of U-2 OS cells at the CEP290 IVS26 locus with two SaCas9 guides ~ 1.1 kb apart generates many possible outcomes measured and depicted in the pie chart. The events include: small indels, the desired ~ 1.1 kb deletion, the ~ 1.1 kb inversion of the intervening fragment, and homologous or sister chromosome translocations. b. A linearity and Lower Limit of Detection (LLoD) experiment using a clonal, engineered HEK293 cell line, mixed at various ratios with the unmodified parental line. The assay has excellent linearity, accuracy, and an LLoD of ~ 0.1%. c. Inter-chromosomal translocations in T-cells are measured after nucleofection with two SpCas9 RNPs, one targeting the TRAC gene and another the B2M gene. The schematic and table shows all possible outcomes and the measured result when applicable. Red lines indicate the editing sites and arrows the primers (1 = OLI6259 and 2 = OLI6256)