Fig. 1.

Genome editing with LbCpf1 in vitro and in vivo. a Mutation frequencies at the Vegfa and Hif1a target sites in C2C12 cells co-transfected with various crRNAs and LbCpf1 genome editing efficiencies were examined by deep sequencing using genome DNA isolated from cells after 48 h of transfection. Error bars indicate s.e.m. (n = 3). b The Cpf1 target sequences in the Vegfa and Hif1a genes. The PAM sequence and the crRNA target sequence (TS3) are shown in blue and red, respectively. c All-in-one AAV vector plasmids encoding Cpf1 and its crRNA (TS3) and its mutation frequencies at the Vegfa and Hif1a target sites in C2C12 cells transfected with pAAV-Cpf1-Vegfa or Hif1a plasmids 48 h post-transfection. d,e Indel frequencies and mutant sequences presented with two fractions, in-frame versus out-of-frame indels at the Vegfa (d) or Hif1a (e) target site in the retina and RPE cells using deep sequencing 6 weeks post-intravitreal injection of AAV-Cpf1-Vegfa or AAV-Cpf1-Hif1a. The PAM sequence and the crRNA target sequence (TS3) are shown in blue and red, respectively. Error bars indicate s.e.m. (n = 4)