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. Author manuscript; available in PMC: 2022 Jul 22.
Published in final edited form as: Angew Chem Int Ed Engl. 2020 Sep 24;59(46):20545–20551. doi: 10.1002/anie.202010648

Figure 3. Two-pair CRISPR processed target gene multiplexed signaling pathway.

Figure 3.

a) Four separate sgRNAs direct four Cas9 D10A nucleases to adjacent target sites, transforming the target into three fragments containing two distinct 3’overhangs accessible for molecular cascading. b) Two orthogonal translator hairpins are initiated by available target sites resulted from CRISPR processing. Target information is translated into an arbitrary sequence, which is further integrated into the same amplifier hairpin, producing a signaling concatemer. c) Comparison of signal gain between single signaling pathway and multiplexed signaling pathway based on different concentrations of gene input. Signal gain %=(peak current-baseline current[without gene condition])/baseline current). d) A dose-dependent electrochemical response of integrated biochemical circuit in a range of concentrations of gene input. The bar represents the mean value of three orthogonal repeats. The error bar represents ± SE. **P<0.05.