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Nature communications
02 07, 2020;11 (1): 779.

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

Patrick S Donahue, Joseph W Draut, Joseph J Muldoon, Hailey I Edelstein, Neda Bagheri, Joshua N Leonard
Author Information
  1. Patrick S Donahue: Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, 60208, USA. ORCID
  2. Joseph W Draut: Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, 60208, USA.
  3. Joseph J Muldoon: Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, 60208, USA. ORCID
  4. Hailey I Edelstein: Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, 60208, USA. ORCID
  5. Neda Bagheri: Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, 60208, USA. ORCID
  6. Joshua N Leonard: Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, 60208, USA. j-leonard@northwestern.edu. ORCID
PMID: 32034124 DOI: 10.1038/s41467-019-14147-5

Abstract

Engineering mammalian cells to carry out sophisticated and customizable genetic programs requires a toolkit of multiple orthogonal and well-characterized transcription factors (TFs). To address this need, we develop the COmposable Mammalian Elements of Transcription (COMET)-an ensemble of TFs and promoters that enable the design and tuning of gene expression to an extent not, to the best of our knowledge, previously possible. COMET currently comprises 44 activating and 12 inhibitory zinc-finger TFs and 83 cognate promoters, combined in a framework that readily accommodates new parts. This system can tune gene expression over three orders of magnitude, provides chemically inducible control of TF activity, and enables single-layer Boolean logic. We also develop a mathematical model that provides mechanistic insights into COMET performance characteristics. Altogether, COMET enables the design and construction of customizable genetic programs in mammalian cells.

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Grants

  1. T32 GM008449/NIGMS NIH HHS
  2. T32 GM008152/NIGMS NIH HHS
  3. T32 GM144295/NIGMS NIH HHS
  4. R01 EB026510/NIBIB NIH HHS
  5. P30 CA060553/NCI NIH HHS
  6. F30 CA203325/NCI NIH HHS

MeSH Term

Animals
Cell Line
Genetic Engineering
HEK293 Cells
Humans
Mammals
Plasmids
Promoter Regions, Genetic
Protein Engineering
Small Molecule Libraries
Transcription Factors
Transcription, Genetic
Zinc Fingers

Chemicals

Small Molecule Libraries
Transcription Factors
Journal Article Research Support, N.I.H., Extramural
Article has an altmetric score of 129

Word Cloud

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