FIGURE 5.

Altering the PAM specificity of eSpCas9PP^D10A and narrowing the editable window of APOBEC1. (A) The eSpCas9pp^D10A–NG gene was cloned into pSEVA6BE, generating pSEVA6BE-NG. (B) C-to-T substitution was successfully achieved in target spacers HexR2 and HexR3 with NG PAM sequences. (C) The APOBEC1 gene in pSEVA6BE was replaced by the APOBEC1-YE1 gene, thus creating pSEVA6BE-YE. (D) pSEVA6BE-YE1-mediated base editing of TtgA-2 spacer tend to have a narrow editing windows compared to using pSEVA6BE (Figure 3F).