Fig. 4
From: Target preference of Type III-A CRISPR-Cas complexes at the transcription bubble
Target preference of TthCsm at the transcription bubble. a Cleavage of nucleic acids by TthCsm in the TthCsm-TEC complex was initiated by addition of MgCl2 or MnCl2 and reactions were incubated for 30 min at 65 ˚C. Products were analyzed by denaturing PAGE and SYBR gold staining. The nontemplate strand (NTS DNA), template strand (TS DNA), RNA transcript (RNA), and crRNAs contained in the TthCsm (Csm crRNAs) were loaded individually on the left half of gel for comparison. b TthCsm-catalyzed cleavage of the 5´-radiolabeled NTS. The substrates tested were the eluted TthCsm-TEC sample from the pull-down (left), the NTS DNA alone with the target RNA transcript, which includes a sequence complementary to the TthCsm crRNA (center), and an R-loop bubble composed of NTS DNA, TS DNA, and the target RNA transcript (right). A ssDNA marker was run in the leftmost lane (M), and the position of the unpaired region is shown. Uncropped gel images for (a, b) are available online in the Source Data file
