Skip to main content
. 2020 Jan 25;48(5):2486–2501. doi: 10.1093/nar/gkz1218

Figure 3.

Figure 3.

Cascade/I-C provides single-stranded DNA loop for Cas3/I-C binding. (A) A schematic model that suggests that Cascade/I-C binding to target DNA provides an ssDNA platform for Cas3 binding. The location of PAM (TTC in red) in the 100 bp construct is indicated. (B) DNA cleavage was tested on 20% native PAGE with increasing concentration of Cas3/I-C in the presence of Cascade (lanes 2–4). Proteinase K was added to release bound DNA (lane 5). No cleavage was noticed in the absence of Cascade (lane 6). (C) A 100 bp bubble DNA construct that lacks base pairing in the center was designed with PAM (TTC in red). The single-stranded DNA loop is suggested to facilitate Cas3/I-C binding in the absence of Cascade/I-C. (D) Bubble DNA was cleaved in the absence of Cascade/I-C (lanes 3–9), whereas dsDNA was cleaved only in the presence of Cascade/I-C (lanes 11–12). DNA was visualized using 20% native PAGE.