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. 2021 Jul 1;49(13):7628–7643. doi: 10.1093/nar/gkab590

Figure 3.

Figure 3.

LdCsm performs indiscriminate DNA degradation at the transcriptional bubble of target and non-target in vitro. Schematic of the Target dsDNA (A) and the Nontarget dsDNA (C). The Target dsDNA substrate contains a T7 promoter and the target sequence (backbone of the protospacer is highlighted yellow and CTR is in red), and the direction of transcription is indicated with an arrow. Lines in the bottom indicate the positions of T7 promoter and target sequence in target dsDNA substrate as well as the size of the full length dsDNAs. The Nontarget dsDNA substrate contains the T7 promoter and the non-homologous sequence, lines in the bottom indicate the position of T7 promoter. Denaturing PAGE analysis of the cleavage products of the Target dsDNA (B) or the Nontarget dsDNA (D). Either non-template strand (Top panel) or template strand (bottom panel) was labeled. Cleavage assay was conducted for 1, 2 and 3 h. Reference reactions lacking one of assay components were conducted for 180 min. M: ssDNA size ladder.