Fig. 2: Comparison of the trans-cleavage activity between Cas12a with WT crRNA and split crRNA (26-nt scaffold + 14-nt spacer).

a Trans-cleavage of ssDNA probes by Cas12a complexed with WT crRNA (blue) and split crRNA (orange) in the presence of target or non-target dsDNA. The reactions contained 50 nM Cas12a, 100 nM of either crRNA or split RNA,10 nM of either complementary dsDNA activators or non-target dsDNA substrates, as well as 20 nM ssDNA probes. b Trans-cleavage of ssDNA probes by complexed with WT crRNA (blue) and split crRNA (orange) in the presence of target or non-target ssDNA. The reactions contained 50 nM Cas12a, 100 nM of either crRNA or split RNA, 10 nM of either complementary ssDNA activators or non-target ssDNA substrates, as well as 20 nM ssDNA probes. c, d Detection of target RNA by Cas12a complexed with scaffold RNA and dsDNA (c) or ssDNA (d) activators. The reaction mixtures were incubated for 60 min at 37 °C and contained 250 nM of Cas12a, 500 nM of scaffold RNA, 10 nM target or non-target RNA substrates, 500 nM of dsDNA or ssDNA activators (dsDNA for c and ssDNA for d), and 1000 nM of fluorescent probes. For all panels, experiments were conducted in triplicate, and error bars represent the mean ± SD (n = 3). Supplementary Data 1 are provided as a Supplementary Data 1 file.