Fig. 4: Programmable transcription activation by Mos350 Cascade-VPR. | Nature Communications

Fig. 4: Programmable transcription activation by Mos350 Cascade-VPR.

From: Engineered minimal type I CRISPR-Cas system for transcriptional activation and base editing in human cells

Fig. 4

a Fold activations of the IL1B gene by Cascades from different strains with Cas7 fused to VPR [P values from left to right: >0.9999, 0.9998, 0.9983, 0.9998, <0.0001]. b Left: Gene architectures of VPR fused with different subunits of the I-F2 Mos350 Cascade. Middle: Cascade with different subunits fused to VPR may result in different locations and copy numbers of VPRs. Orange: Cas7; Yellow: Cas6; Cyan: Cas5; Blue: VPR. Right: Fold activations by different VPR fusion approaches [P values from left to right: 0.0632, 0.9346, <0.0001]. c Fold activations of three genes (ACTC1, ASCL1, and NEUROD1) when the I-F2 Mos350 Cascade-VPR (Cas7-VPR) targets different regions upstream of the TSS [P values from left to right: 0.0207, <0.0001, 0.0058, 0.0002, 0.0002, 0.0003; 0.9997, <0.0001, 0.4304, >0.9999, 0.9997, 0.8879; <0.0001, <0.0001, 0.6434, 0.0007, 0.9997, 0.9868]. d Histograms showing normalized mean transcriptional activation levels of ACTC1, ASCL1, and NEUROD1, targeting different regions upstream of the TSS. The data normalized are from (c). Normalized values for all 3 genes are plotted as box-and-whisker plots with min to max as an option. Maximum and minimum values are shown as whiskers. The boxes in the graph display the median value and the lower and upper quartiles. e Fold activations of the HBB and HBG1 genes with I-F2 Mos350 Cascade-VPR (Cas7-VPR) targeting 0 to 200 bp upstream of the TSS [P values from left to right: <0.0001, <0.0001; <0.0001, 0.5049]. In (a) and (b), ctrl: None plasmid was transformed. In (c) and (e), ctrl: Only the plasmid expressing the Cascade-VPR was transformed. All values are represented as mean ± s.d. (n = 3 biological replicates). Internal reference gene (a–c) and (e): GAPDH. Statistical significance was assessed by one-way ANOVA with Dunnett’s multiple comparisons tests (*P < 0.05; **P < 0.01; ***P < 0.001). In (a–c) and (e), each bar chart shows the results of samples treated in the same batch. Each experiment was conducted at least in two batches with similar differential trends. Source data are provided as a Source data file.

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