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The COMET toolkit for composing customizable genetic programs in mammalian cells.

Patrick S Donahue1,2,3, Joseph W Draut1, Joseph J Muldoon1,2

  • 1Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, 60208, USA.

Nature Communications
|February 9, 2020
PubMed
Summary
This summary is machine-generated.

Researchers developed the COmposable Mammalian Elements of Transcription (COMET) system for engineering mammalian cells. This toolkit enables precise control over gene expression, facilitating the design of complex genetic programs.

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Area of Science:

  • Synthetic biology
  • Molecular and cell biology
  • Genetic engineering

Background:

  • Engineering mammalian cells for complex genetic programs requires orthogonal transcription factors (TFs).
  • Existing toolkits may lack the necessary components for sophisticated gene expression control.

Purpose of the Study:

  • To develop a comprehensive and customizable toolkit for designing and tuning gene expression in mammalian cells.
  • To create an ensemble of transcription factors and promoters for advanced genetic engineering.

Main Methods:

  • Developed the COmposable Mammalian Elements of Transcription (COMET) system.
  • Integrated 44 activating and 12 inhibitory zinc-finger TFs with 83 cognate promoters.
  • Incorporated chemically inducible control and a framework for new part integration.
  • Created a mathematical model for mechanistic insights into system performance.

Main Results:

  • COMET enables gene expression tuning over three orders of magnitude.
  • The system provides chemically inducible control of TF activity.
  • COMET supports single-layer Boolean logic operations.
  • A mathematical model elucidated COMET's performance characteristics.

Conclusions:

  • COMET provides an unprecedented level of control for designing and constructing customizable genetic programs in mammalian cells.
  • The toolkit's modularity and extensive component library facilitate advanced synthetic biology applications.