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Tunable Temperature-Sensitive Transcriptional Activation Based on Lambda Repressor.

Lealia L Xiong1, Michael A Garrett2, Marjorie T Buss2

  • 1Division of Engineering and Applied Sciences, California Institute of Technology, 1200 E. California Blvd., Pasadena, California 91125, United States.

ACS Synthetic Biology
|June 16, 2022
PubMed
Summary
This summary is machine-generated.

Researchers developed novel cold-activated gene switches using temperature-sensitive Lambda phage repressor CI variants. These engineered transactivators enable tunable, cooling-induced gene expression in E. coli, expanding synthetic biology tools.

Keywords:
microbial synthetic biologytemperaturethermal controltransactivationtranscription factors

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Genetic Engineering

Background:

  • Temperature control is crucial for engineered cellular functions.
  • Heat-inducible gene expression is well-established using temperature-sensitive repressors.
  • Few cold-inducible transcriptional activators exist, limiting temperature-based cellular control.

Purpose of the Study:

  • To investigate temperature-dependent Lambda phage repressor CI mutants as cold-actuated transactivators.
  • To engineer tunable, cooling-induced gene expression systems.
  • To expand the toolbox for cellular function control using thermal inputs.

Main Methods:

  • Utilized thermolabile mutants of Lambda phage repressor CI (TcI family).
  • Tested TcI variants for temperature-sensitive transcriptional activation at the PRM promoter in E. coli.
  • Demonstrated simultaneous activation and repression of different genes within the same circuit.

Main Results:

  • Two TcI variants showed temperature-responsive activation with set points at 35.5 °C and 38.5 °C.
  • Successfully created cold-inducible operons with tunable temperature responses.
  • TcI demonstrated dual functionality as both an activator and repressor, exhibiting opposite thermal responses.

Conclusions:

  • Temperature-dependent Lambda phage repressor CI variants function as tunable cold-actuated transactivators.
  • This expands the available tools for precise temperature-mediated control of gene expression in synthetic biology.
  • The dual activator/repressor capability offers advanced circuit design possibilities for cellular control.