2022
DOI: 10.1021/acssynbio.1c00545
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Highly Reversible Tunable Thermal-Repressible Split-T7 RNA Polymerases (Thermal-T7RNAPs) for Dynamic Gene Regulation

Abstract: Temperature is a physical cue that is easy to apply, allowing cellular behaviors to be controlled in a contactless and dynamic manner via heat-inducible/repressible systems. However, existing heat-repressible systems are limited in number, rely on thermal sensitive mRNA or transcription factors that function at low temperatures, lack tunability, suffer delays, and are overly complex. To provide an alternative mode of thermal regulation, we developed a library of compact, reversible, and tunable thermalrepressi… Show more

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Cited by 23 publications
(17 citation statements)
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“…Despite their favorable features, temperature-responsive genetic circuits still present some limitations. One of the critical challenges is to modify the dynamic range, and to address this, the effects of the CI 857 expression on the biosensor response should be explored for in future works ( 36 ). Additionally, the performances of genetic circuits are highly dependent on the physiological conditions of the cells.…”
Section: Discussionmentioning
confidence: 99%
“…Despite their favorable features, temperature-responsive genetic circuits still present some limitations. One of the critical challenges is to modify the dynamic range, and to address this, the effects of the CI 857 expression on the biosensor response should be explored for in future works ( 36 ). Additionally, the performances of genetic circuits are highly dependent on the physiological conditions of the cells.…”
Section: Discussionmentioning
confidence: 99%
“…As an alternative to pure RNA modules, RNA and protein thermosensors can be combined to amplify the the temperature response. For this purpose, the temperature-sensitive coiled-coil domain of the Tlpa protein can be fused to the protein of interest [130] , [131] .…”
Section: Biotechnological Applications Of Temperature Controlmentioning
confidence: 99%
“…Recently, the Poh research group reported a reversibly controlled temperature-sensitive gene expression system by the fusion of the C-terminal coiled-coil domain of TlpA (Ala94–Ala371 region, 278 amino acids, TlpA coil) with split-T7 RNAP. To generate the Thermal T7RNAP, N-terminal fragment of T7 RNAP (Met1–Ser564)–TlpA coil and TlpA coil–C-terminal fragment of T7 RNAP (Glu565–Ala883) fusions were used [ 14 ] ( Figure 2 B).…”
Section: Temperature-controlled Gene Expressionmentioning
confidence: 99%
“…From a synthetic biology perspective, nature’s mechanisms for sensing temperature and light have provided powerful insights for designing tools to achieve conditional gene expression. At the molecular level, researchers have utilized the heat shock response mechanism to construct temperature-sensitive gene expression systems such as the temperature-sensitive dimeric repressor [ 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 ], RNA polymerase (RNAP) [ 14 , 15 , 16 , 17 ], replication machinery [ 18 , 19 , 20 , 21 , 22 ], and oligonucleotide topology [ 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 ]. Photocaged [ 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 ] or photoswitchable [ 44 , 45 ] small-molecule inducers and photocaged transcription factors [ 46 ] have been used to develop light-driven gene expression systems.…”
Section: Introductionmentioning
confidence: 99%