Fig. 6: Design of PRCIS-based orthogonal reporting systems and logic gates. | Nature Communications

Fig. 6: Design of PRCIS-based orthogonal reporting systems and logic gates.

From: A generalizable approach for programming protease-responsive conformationally inhibited artificial transcriptional factors

Fig. 6

Performance of (a) eGCN+/+ and (b) eMAX+/+ with different PCP linkers upon corresponding protease activation. c Color map summerizes fold changes of PRCIS equipped with different ATFs and PCPs in respond to corresponding proteases. d Schematic illustration and performance of the tri-orthogonal PRCIS-based circuit consisting of TVMVp-inducible eGCN+/+ (eGCN+/+(TVMVpcp)), PPVp-inducible eGAL+/+ (eGAL+/+(PPVpcp)) and TEVp-inducible eMAX+/+ (eAMX+/+(TEVpcp)) for the reporting of all combinations of TEVp, TVMVp, and PPVp inputs. e Schematic illustration and evaluation of orthogonal reporting of ABA and RAP with ABA-inducible TEVp-responsive eGCN+/+ (eGCN+/+(TEVpcp)) and RAP-inducible TVMVp-responsive eGAL+/+ (eGAL+/+(TVMVpcp)). Schematic illustration and evaluation of PRCIS-based (f) ‘OR’ gate, (g) ‘AND’ gate, and (h) ‘INHIBIT’ gate with TEVp and TVMVp inputs and mCherry outputs. n = 3, individual data points represent independent triplicates performed on the same day. For (d, e), error bars represent MFI (arbs.units) ± SD, For (a, b, f, g, h), error bars represent MMFI (arbs.units) ± SD. Significance (two-tailed, unpaired t test) is labeled above data, ns: not significant, **p < 0.01, ***p < 0.001, ****p < 0.0001. Schematic illustrations of (d, e, f, g, h): Created in BioRender. He, J. (2025) https://BioRender.com/t4qifbc. Source data are provided as a Source Data file.

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