Synthetic logic circuits using RNA aptamer against T7 RNA polymerase
| Autor: | Juan F Quijano, Jongmin Kim, Richard M. Murray, Enoch Yeung, Jeongwon Kim |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
0106 biological sciences
Logic Aptamer Cell Computational biology 01 natural sciences Applied Microbiology and Biotechnology Metabolic engineering Synthetic biology Viral Proteins Transcription (biology) 010608 biotechnology medicine Escherichia coli T7 RNA polymerase Chemistry 010401 analytical chemistry RNA General Medicine DNA-Directed RNA Polymerases Aptamers Nucleotide 0104 chemical sciences medicine.anatomical_structure Nucleic acid Molecular Medicine Synthetic Biology medicine.drug |
| Popis: | Recent advances in nucleic acids engineering introduced several RNA-based regulatory components for synthetic gene circuits, expanding the toolsets to engineer organisms. In this work, we designed genetic circuits implementing an RNA aptamer previously described to have the capability of binding to the T7 RNA polymerase and inhibiting its activity in vitro. Using in vitro transcription assays, we first demonstrated the utility of the RNA aptamer in combination with programmable synthetic transcription networks. As a step to quickly assess the feasibility of aptamer functions in vivo, a cell-free expression system was used as a breadboard to emulate the in vivo conditions of E. coli. We tested the aptamer and its three sequence variants in the cell-free expression system, verifying the aptamer functionality in the cell-free testbed. In vivo expression of aptamer and its variants demonstrated control over GFP expression driven by T7 RNA polymerase with different response curves, indicating its ability to serve as building blocks for both logic circuits and transcriptional cascades. This work elucidates the potential of RNA-based regulators for cell programming with improved controllability leveraging the fast production and degradation time scales of RNA molecules. |
| Databáze: | OpenAIRE |
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