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Synthetic logic circuits using RNA aptamer against T7 RNA polymerase

Kim, Jongmin and Quijano, Juan F. and Kim, Jeongwon and Yeung, Enoch and Murray, Richard M. (2021) Synthetic logic circuits using RNA aptamer against T7 RNA polymerase. Biotechnology Journal . ISSN 1860-7314. doi:10.1002/biot.202000449. (In Press)

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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. We first demonstrated the utility of the RNA aptamer in combination with programmable synthetic transcription networks in vitro. As a step to quickly assess the feasibility of aptamer functions in vivo, we tested the aptamer and its sequence variants in the cell‐free expression system, verifying the aptamer functionality in the cell‐free testbed. The expression of aptamer in E. coli demonstrated control over GFP expression driven by T7 RNA polymerase, indicating its ability to serve as building blocks for logic circuits and transcriptional cascades. This work elucidates the potential of T7 RNA polymerase aptamer as regulators for synthetic biological circuits and metabolic engineering.

Item Type:Article
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URLURL TypeDescription Paper
Kim, Jongmin0000-0002-2713-1006
Yeung, Enoch0000-0001-7630-7429
Murray, Richard M.0000-0002-5785-7481
Additional Information:© 2021 Wiley. Accepted manuscript online: 04 April 2021; Manuscript accepted: 30 March 2021; Manuscript revised: 05 March 2021; Manuscript received: 21 October 2020. This work was supported by National Science Foundation award no. 0832824 (The Molecular Programming Project) and the Defense Advanced Research Projects Agency (DARPA/MTO) Living Foundries program, contract number HR0011-12-C-0065 (DARPA/CMO). The views and conclusions contained in this document are those of the authors and should not be interpreted as representing officially policies, either expressly or implied, of the Defense Advanced Research Projects Agency or the U.S. Government. The authors thank David Shis and Matthew Bennett for providing the pTara plasmid. Conflict of interest statement. None declared.
Funding AgencyGrant Number
Defense Advanced Research Projects Agency (DARPA)HR0011-12-C-0065
Subject Keywords:aptamers; cell‐free expression systems; genelet; metabolic engineering; synthetic biology
Record Number:CaltechAUTHORS:20140925-085129247
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Official Citation:Kim, J., Quijano, J.F., Kim, J., Yeung, E. and Murray, R.M. (2021), Synthetic logic circuits using RNA aptamer against T7 RNA polymerase. Biotechnol. J. Accepted Author Manuscript 2000449.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:50017
Deposited By: Jason Perez
Deposited On:14 May 2016 03:27
Last Modified:08 Apr 2021 22:14

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