Multi-Ribozyme-gRNA-Aptazyme (RGA) networks for biosensing and gene regulation
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Biological parts can be considered as molecular building blocks to assemble complex synthetic genetic networks utilizing synthetic biology. In this work, we constructed three types of multi-Ribozyme-gRNA-Aptazyme (RGA) networks for biosensing and gene regulation by linking RGA, Ribozyme-gRNA-Ribozyme (RGR) and CRISPRi systems together. We screened out two different aptazymes that fit this network and analyzed their one-layer and two-layer single-input RGAs behaviors and functions in two-inputs systems with different architectures. Overall, we built two IMPLY gates, two NIMPLY gates, one AND gate, one partial NAND gate, one conditional non-Boolean gate. The RGA shows as much as a 7.38-fold increase in ligand-dependent system-output. The interaction between RGAs that are inserted in circuits in parallel has also been reported, which contributions to the predictability of more complicated RGA networks.
- Chemical engineering