Fig. 4: Multiplexed RNA degradation using dCasCMA and multiplexed gRNA expression array.
From: Programmable targeted RNA degradation via dCas13d-directed chaperone-mediated autophagy (dCasCMA)
a Schematic representation of the ZBP1 and STING signaling pathways activated by viral infection. Created in BioRender. Wen, H. (2025) https://BioRender.com/41b43vf. b Schematic showing the design of multiplexed gRNA expression arrays (gJZS) targeting JEV RNA, ZBP1 mRNA, and STING mRNA using the dCasCMA system. dCCTM processes the gRNA arrays into mature gRNAs for targeted RNA degradation. Created in BioRender. Wen, H. (2025) https://BioRender.com/41b43vf. c RNA expression levels in infected cells treated with dCCTM/gJZS. (vRNA p < 0.0001; ZBP1 p = 0.0008; STING p = 0.0001). d, e WB and quantitative analysis of protein expression levels of JEV E, ZBP1 and STING in infected cells treated with dCCTM/gJZS. (JEV E p = 0.0006; ZBP1 p = 0.006; STING p = 0.0002). f Downstream effects of the degradation of JEV RNA, ZBP1 and STING in infected cells treated with dCCTM/gJZS. The expression of pro-inflammatory cytokines and chemokines (TNF-α, IFN-γ, IL-6, IL-1β, IL-18, CCL2, CCL4, CXCL9, and CXCL10) was significantly reduced, while the expression of the anti-inflammatory cytokine Il-10 was significantly increased. dCCTM/NT vs. dCCTM/gJZS (TNF-α p < 0.0001; IFN-γ p < 0.0001; IL-6 p < 0.0001; IL-1β p = 0.0002; IL-10 p = 0.0002; IL-18 p = 0.001; CCL2 p < 0.0001; CCL4 p < 0.0001; CXCL9 p = 0.002; CXCL10 p = 0.0003). Data are presented as means ± SD (n = 3, biological replicates). Statistical significance was determined by unpaired two-tailed Student’s t-test. **p < 0.01 and ***p < 0.001. Source data are provided as a Source Data file.
