Figure 1.

gRNA Design and Validation for the SHOX2 c.849C>A and the SHOX2 c.^∗28T>C Loci

(A) Experimental overview of gRNA validation. hiPSCs were transfected with Cas9 RNP/gRNA complexes and ssODNs. After 48 h, the Cas9-targeted region was amplified and analyzed via NGS.

(B) Boxes, top: The SHOX2 c.849C>A and SHOX2 c.^∗28T>C loci with selected gRNA binding sites. PAM sequences are depicted in blue, the mutated base pair is in red. Boxes, bottom: Indel frequency for each gRNA in hiPSCs 48 h after transfection. The targeted region is centered. The cumulative frequency of deletions (blue) and insertions (green) at each position is depicted in percentage of reads (% Reads). Abbreviations: RNP, ribonucleoprotein; gRNA, single guide RNA; ssODN, single-stranded oligodesoxynucleotide; NGS, next-generation sequencing.

See also Figure S1, Table S1, S2, S5, and S6.