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. Author manuscript; available in PMC: 2018 Aug 30.
Published in final edited form as: J Am Chem Soc. 2017 Aug 15;139(34):11964–11972. doi: 10.1021/jacs.7b06152

Figure 6.

Figure 6

Schematic of a system to detect 4 multidimensional PPIs simultaneously. (A) Design of mammalian vector system to measure a one-by-four PPI network. Expression vectors for a series of anti-apoptotic-RNAPC using the orthogonal RNAPC variants developed in Figure 5 were engineered with a gene circuit that produces an orthogonal RNA output signal from transcription on the orthogonal DNA promoter. Four orthogonal vectors were created, which can then be deployed along with a BH3-only-RNAPN expression vector. (B) Schematic of one-by-four RNAP tagging system for multidimensional PPI analysis. The BH3-only peptide fusion on the RNAPN tag can competitively bind to a series of Bcl-2-family proteins, each themselves fused to orthogonal RNAPC tags. Interactions between each Bcl-2-family protein with the target results in the assembly of an orthogonal RNAP, which in turn transcribes an output RNA signal from an orthogonal DNA promoter. The relative amount of each PPI can be measured by RT-qPCR analysis of the output RNA signals.