Figure 3.

GFP disruption mediated by different Cpf1 nucleases on targets with TTTV PAMs in HEK-293.GFP cells. GFP disruption mediated by different Cpf1 nucleases (blue bars: LbCpf1, orange bars: AsCpf1, green bars: FnCpf1 and yellow bars: MbCpf1) on targets with a TTTA (GFP target 1) or a TTTG (GFP target 2) PAM sequence. The nuclease vector along with the corresponding crRNA expressing vector were transfected into HEK-293.GFP cells and GFP positive cells were counted 7 days post-transfection by flow cytometry. For controls we used either inactive Cpf1 expressing nuclease vectors or no-target crRNA expressing vectors. The average GFP % of negative controls was 95.72 ± 1.13%. GFP disruption activities of the nucleases were calculated as follows: 1 – (sample GFP %/average GFP % of negative controls). Bars correspond to averages of n = 3 parallel samples. The experiment was repeated six times and a representative dataset is presented here.