Fig. 1

DNA cleavage of Cas12a orthologs in vitro. a DNA cleavage activities of BfCas12a and Cas12a in vitro. The Cas12a-crRNAcomplex (100 nM) was incubated at 37 °C for 10 min with DNA substrates (see also Additional file 1: Figure S4). b Schematic of in vitro cleavage assay used to identify PAM sequence and determine global PAM specificity. c Web logo for the CeCas12a PAM. d Web logo for the BfCas12a PAM. e Normalized cleavage rates for all 4-base PAMs for CeCas12a, BfCas12a, LbCas12a, and AsCas12a. The intensity of color represented the activity of Cas12a nuclease. f Quantification of time-course in vitro cleavage reactions of Cas12a orthologs on linearized dsDNA substrates. Those were conducted at nine time points, respectively. Curves were fit using the one phase exponential decay equation