Supplementary Figure 1: Activities of Cas12a orthologs and engineered variants in human cells. | Nature Biotechnology

Supplementary Figure 1: Activities of Cas12a orthologs and engineered variants in human cells.

From: Engineered CRISPR–Cas12a variants with increased activities and improved targeting ranges for gene, epigenetic and base editing

Supplementary Figure 1

(a) Activities of Cas12a orthologs targeted to endogenous sites in human cells bearing TTTN or VTTN PAMs. Percent modification assessed by T7E1 assay; mean, s.e.m., and individual data points shown for n = 3. (b) Summary of the activities of Cas12a orthologs against 24 sites with NTTN PAM sequences (mean activities shown from data in panel a). *, P < 0.05; ****, P < 0.0001 (Mann–Whitney, two-tailed; P values in Supplementary Table 8). (c) Schematic and structural representations of Cas12a paired with a crRNA, and interacting with a target site encoding a prototypical TTTA PAM. In structural representations, amino acid residues proximal to PAM DNA bases are highlighted in green; images generated from PDBID:5B43 (ref. 17) visualized in PyMOL (v 1.8.6.0); select regions of the PAM-interacting domain are hidden for clarity. (d, e) Activities of AsCas12a variants bearing single amino acid substitutions when tested against endogenous sites in human cells bearing canonical (panel d) or non-canonical (panel e) PAMs. Percent modification assessed by T7E1 assay; mean, s.e.m., and individual data points shown for n = 3. (f) Modification of endogenous sites in human cells by wild-type AsCas12a and variants bearing amino acid substitutions. Activities assessed by T7E1 assay; mean, s.e.m., and individual data points shown for n = 3. Reference. 17. Yamano, T. et al. Crystal Structure of Cpf1 in Complex with Guide RNA and Target DNA. Cell 165, 949–962 (2016).

Back to article page