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Single-cell measurement of plasmid copy number and promoter activity

Author

Listed:
  • Bin Shao

    (Massachusetts Institute of Technology)

  • Jayan Rammohan

    (National Institute of Standards and Technology)

  • Daniel A. Anderson

    (Massachusetts Institute of Technology)

  • Nina Alperovich

    (National Institute of Standards and Technology)

  • David Ross

    (National Institute of Standards and Technology)

  • Christopher A. Voigt

    (Massachusetts Institute of Technology)

Abstract

Accurate measurements of promoter activities are crucial for predictably building genetic systems. Here we report a method to simultaneously count plasmid DNA, RNA transcripts, and protein expression in single living bacteria. From these data, the activity of a promoter in units of RNAP/s can be inferred. This work facilitates the reporting of promoters in absolute units, the variability in their activity across a population, and their quantitative toll on cellular resources, all of which provide critical insights for cellular engineering.

Suggested Citation

  • Bin Shao & Jayan Rammohan & Daniel A. Anderson & Nina Alperovich & David Ross & Christopher A. Voigt, 2021. "Single-cell measurement of plasmid copy number and promoter activity," Nature Communications, Nature, vol. 12(1), pages 1-9, December.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-21734-y
    DOI: 10.1038/s41467-021-21734-y
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    Cited by:

    1. Carolyn N. Bayer & Maja Rennig & Anja K. Ehrmann & Morten H. H. Nørholm, 2021. "A standardized genome architecture for bacterial synthetic biology (SEGA)," Nature Communications, Nature, vol. 12(1), pages 1-13, December.
    2. Miles V. Rouches & Yasu Xu & Louis Brian Georges Cortes & Guillaume Lambert, 2022. "A plasmid system with tunable copy number," Nature Communications, Nature, vol. 13(1), pages 1-12, December.
    3. John P. Marken & Richard M. Murray, 2023. "Addressable and adaptable intercellular communication via DNA messaging," Nature Communications, Nature, vol. 14(1), pages 1-11, December.
    4. Michael B. Sheets & Nathan Tague & Mary J. Dunlop, 2023. "An optogenetic toolkit for light-inducible antibiotic resistance," Nature Communications, Nature, vol. 14(1), pages 1-13, December.
    5. J. Carlos R. Hernandez-Beltran & Jerónimo Rodríguez-Beltrán & Oscar Bruno Aguilar-Luviano & Jesús Velez-Santiago & Octavio Mondragón-Palomino & R. Craig MacLean & Ayari Fuentes-Hernández & Alvaro San , 2024. "Plasmid-mediated phenotypic noise leads to transient antibiotic resistance in bacteria," Nature Communications, Nature, vol. 15(1), pages 1-13, December.

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