Additional file 1: of All-in-one adeno-associated virus delivery and genome editing by Neisseria meningitidis Cas9 in vivo

Figure S1. Genome-wide computational prediction of NmeCas9 off-target sites using CRISPRseek with the N4GN3 PAM. Search parameters were set to identify sites with up to six mismatches to the spacer sequence. The total number of detected off-target sites for each protospacer is indicated. Figure S2. Stacked histogram showing the percentage distribution of indels obtained by TIDE after AAV-sgRNA-hNmeCas9 plasmids transfections in Hepa1–6. Error bars represent three independent experiments. Data are presented as mean ± SD. Figure S3. H&E staining from wild-type (Fah+/+) mouse, and HT-I mice (Fah−/−) injected with PBS or AAV-sgRNA-hNmeCas9 plasmids sgHpd1 or sgHpd2. Scale bar is 20 μm. Figure S4. Humoral IgG1 immune response to NmeCas9 in vivo. Serum collected at day 50 post injection with all-in-one AAV8-sgRNA-hNmeCas9 sgPcsk9 and sgRosa26. Serum antibodies reacted against NmeCas9 protein (left) or SpyCas9 protein (right). Figure S5. H&E staining from PBS, sgPcsk9, and sgRosa26 AAV8 injected mice. Scale bar is 20 μm. (PDF 713 kb