Fig. 2: Investigation of CBS features for EcCas6 cleavage. | Nature Communications

Fig. 2: Investigation of CBS features for EcCas6 cleavage.

From: De novo design of hypercompact transcript degraders by engineering substrate-specific toxins and Cas6-CBS system

Fig. 2

a Schematic of four distinct regions of wtCBS, with numbered bases. The cleavage site (marked by a black triangle) indicates the GA junction between the stem and the 3′ flanking sequence cleaved by EcCas6. b Schematic of the enCBS-GFP-OFF reporter system, designed to assess the functionality of various CBS sequences in response to EcCas6 cleavage activity. c Investigation of the impact of stem truncation and extension of CBS on EcCas6 cleavage activity. d Investigation of the impact of the loop size of CBS with varying stem lengths on EcCas6 cleavage activity. e Investigation of the impact of combination engineering in the four regions of the wtCBS on EcCas6 cleavage activity. Negative control (NC) represents the GFP reporter plasmid without enCBS. GFP fluorescence values are normalized to the dEcCas6 control. Details of the tested enCBS variants are available in Supplementary Data 3. Data are mean ± s.e.m. of n = 3 biological replicates. Source data are provided as a Source data file.

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