Fig. 3: Investigation of CBS features for EcCas6 and dEcCas6 binding. | Nature Communications

Fig. 3: Investigation of CBS features for EcCas6 and dEcCas6 binding.

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

Fig. 3

a Schematic of the EcCas6-VPR-based CRISPRa reporter system in response to EcCas6 binding affinity. b Left: Schematic of the dSpyCas9-VPR-based CRISPRa system with sgRNA-enCBS. Right: Fold increase in fluorescence for the seven sgRNA-enCBS variants and sgRNA in the dSpyCas9-VPR-based CRISPRa system, compared with the control expressing dSpyCas9-VPR and non-targeting sgRNA. c Fold increase in fluorescence for the seven sgRNA-enCBS variants induced by EcCas6-VPR or dEcCas6-VPR, compared with the control expressing dSpyCas9, non-targeting sgRNA-wtCBS, and EcCas6-VPR or dEcCas6-VPR. d Schematic of the working principle of the enCBS-GFP-OFF reporter system using dEcCas6-barnase 11. e Comparison of the degradation efficiency of a panel of enCBS-GFP mRNAs, induced by dEcCas6-barnase 11 and EcCas6. GFP fluorescence values are normalized to the dEcCas6 control. Data are mean ± s.e.m. of n = 3 biological replicates. Source data are provided as a Source data file.

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