Figure 5.

Reverse Genome Editing to Determine the Initial HDR Efficiency

(A) Experimental scheme for reverse gene editing. The newly generated isogenic clones were used to determine the initial HDR efficiency. For this, the same Cas9 RNP and gRNA were used in combination with an oligonucleotide donor differing only in the one base analogous to SHOX2 c.849 (B) or SHOX2 c.^∗28 (C). In consequence, HDR events would lead to the introduction of the SHOX2 c.849C>A or SHOX2 c.^∗28T>C mutation. The frequency of HDR was determined by NGS 72 h after isogenic cells were transfected with Cas9/gRNA/ssODNs. Abbreviations: HDR, homology-directed repair.

See also Tables S2, S5, and S6.