Engineered systems of inducible anti-repressors for the next generation of biological programming

Thomas M. Groseclose, Ronald E. Rondon, Zachary D. Herde, Carlos A. Aldrete and Corey J. Wilson ()
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Thomas M. Groseclose: School of Chemical & Biomolecular Engineering
Ronald E. Rondon: School of Chemical & Biomolecular Engineering
Zachary D. Herde: School of Chemical & Biomolecular Engineering
Carlos A. Aldrete: School of Chemical & Biomolecular Engineering
Corey J. Wilson: School of Chemical & Biomolecular Engineering

Nature Communications, 2020, vol. 11, issue 1, 1-15

Abstract: Abstract Traditionally engineered genetic circuits have almost exclusively used naturally occurring transcriptional repressors. Recently, non-natural transcription factors (repressors) have been engineered and employed in synthetic biology with great success. However, transcriptional anti-repressors have largely been absent with regard to the regulation of genes in engineered genetic circuits. Here, we present a workflow for engineering systems of non-natural anti-repressors. In this study, we create 41 inducible anti-repressors. This collection of transcription factors respond to two distinct ligands, fructose (anti-FruR) or D-ribose (anti-RbsR); and were complemented by 14 additional engineered anti-repressors that respond to the ligand isopropyl β-d-1-thiogalactopyranoside (anti-LacI). In turn, we use this collection of anti-repressors and complementary genetic architectures to confer logical control over gene expression. Here, we achieved all NOT oriented logical controls (i.e., NOT, NOR, NAND, and XNOR). The engineered transcription factors and corresponding series, parallel, and series-parallel genetic architectures represent a nascent anti-repressor based transcriptional programming structure.

Date: 2020
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DOI: 10.1038/s41467-020-18302-1

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