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A synthetic transcription platform for programmable gene expression in mammalian cells

Author

Listed:
  • William C. W. Chen

    (Massachusetts Institute of Technology
    Massachusetts Institute of Technology
    Massachusetts Institute of Technology
    Massachusetts General Hospital)

  • Leonid Gaidukov

    (Massachusetts Institute of Technology
    Massachusetts Institute of Technology)

  • Yong Lai

    (Massachusetts Institute of Technology
    Massachusetts Institute of Technology)

  • Ming-Ru Wu

    (Massachusetts Institute of Technology
    Dana-Farber Cancer Institute and Harvard Medical School)

  • Jicong Cao

    (Massachusetts Institute of Technology)

  • Michael J. Gutbrod

    (Massachusetts Institute of Technology
    Broad Institute of Massachusetts Institute of Technology and Harvard University)

  • Gigi C. G. Choi

    (Massachusetts Institute of Technology
    The University of Hong Kong)

  • Rachel P. Utomo

    (Massachusetts Institute of Technology
    Wellesley College)

  • Ying-Chou Chen

    (Massachusetts Institute of Technology
    Massachusetts Institute of Technology
    National Yang-Ming University)

  • Liliana Wroblewska

    (Pfizer Inc.)

  • Manolis Kellis

    (Massachusetts Institute of Technology
    Broad Institute of Massachusetts Institute of Technology and Harvard University)

  • Lin Zhang

    (Pfizer Inc.)

  • Ron Weiss

    (Massachusetts Institute of Technology
    Massachusetts Institute of Technology)

  • Timothy K. Lu

    (Massachusetts Institute of Technology
    Massachusetts Institute of Technology
    Massachusetts Institute of Technology)

Abstract

Precise, scalable, and sustainable control of genetic and cellular activities in mammalian cells is key to developing precision therapeutics and smart biomanufacturing. Here we create a highly tunable, modular, versatile CRISPR-based synthetic transcription system for the programmable control of gene expression and cellular phenotypes in mammalian cells. Genetic circuits consisting of well-characterized libraries of guide RNAs, binding motifs of synthetic operators, transcriptional activators, and additional genetic regulatory elements express mammalian genes in a highly predictable and tunable manner. We demonstrate the programmable control of reporter genes episomally and chromosomally, with up to 25-fold more activity than seen with the EF1α promoter, in multiple cell types. We use these circuits to program the secretion of human monoclonal antibodies and to control T-cell effector function marked by interferon-γ production. Antibody titers and interferon-γ concentrations significantly correlate with synthetic promoter strengths, providing a platform for programming gene expression and cellular function in diverse applications.

Suggested Citation

  • William C. W. Chen & Leonid Gaidukov & Yong Lai & Ming-Ru Wu & Jicong Cao & Michael J. Gutbrod & Gigi C. G. Choi & Rachel P. Utomo & Ying-Chou Chen & Liliana Wroblewska & Manolis Kellis & Lin Zhang & , 2022. "A synthetic transcription platform for programmable gene expression in mammalian cells," Nature Communications, Nature, vol. 13(1), pages 1-16, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-33287-9
    DOI: 10.1038/s41467-022-33287-9
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    References listed on IDEAS

    as
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