Fig. 6

Genome specificity of IL2RG sgRNA guide. a Heat map of on-target INDEL frequencies quantied by NexGen-Seq at COSMID identified putative on-target locations from healthy CD34⁺ HSPCs. Levels of NHEJ induced by 20 nt IL2RG sgRNA and truncated 19 nt, 18 nt and 17 nt pre-complexed with WT Cas9 protein at 5:1 molar ratio. b Heat map as in (a) of on-target INDEL frequencies derived from 19 nt IL2RG sg-1 in the genome of CD34⁺ HSPCs SCID-X1 patient 1 derived cells. c Percent viability at day 4 of SCID-X1 patient-derived CD34⁺ HSPCs nucleofected with either wild-type (WT) or high-fidelity (HiFi) SpCas9 protein pre-complexed with either the 20 nt or the 19 nt IL2RG sg-1 (n = 1). d Percent INDELs measured by TIDE at day 4 in cells as in (c) using WT or HiFi Cas9 protein pre-complexed with the 20 nt IL2RG sg-1 (green bars) or 19 nt IL2RG sg-1 (blue bars). e Percent IL2RG cDNA targeting (% HR) as measured by ddPCR at day 4 in cells as in (c) generated by either WT or HiFi Cas9 protein pre-complexed with the 20 nt IL2RG sg-1 or (f) 19 nt IL2RG sg-1. Source data are available in the Source Data file