Single-cell measurement of plasmid copy number and promoter activity

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 inf...

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Autores principales: Shao, Bin, Rammohan, Jayan, Anderson, Daniel A., Alperovich, Nina, Ross, David, Voigt, Christopher A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7935883/
https://www.ncbi.nlm.nih.gov/pubmed/33674569
http://dx.doi.org/10.1038/s41467-021-21734-y
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author Shao, Bin
Rammohan, Jayan
Anderson, Daniel A.
Alperovich, Nina
Ross, David
Voigt, Christopher A.
author_facet Shao, Bin
Rammohan, Jayan
Anderson, Daniel A.
Alperovich, Nina
Ross, David
Voigt, Christopher A.
author_sort Shao, Bin
collection PubMed
description 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.
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spelling pubmed-79358832021-03-21 Single-cell measurement of plasmid copy number and promoter activity Shao, Bin Rammohan, Jayan Anderson, Daniel A. Alperovich, Nina Ross, David Voigt, Christopher A. Nat Commun Article 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. Nature Publishing Group UK 2021-03-05 /pmc/articles/PMC7935883/ /pubmed/33674569 http://dx.doi.org/10.1038/s41467-021-21734-y Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Shao, Bin
Rammohan, Jayan
Anderson, Daniel A.
Alperovich, Nina
Ross, David
Voigt, Christopher A.
Single-cell measurement of plasmid copy number and promoter activity
title Single-cell measurement of plasmid copy number and promoter activity
title_full Single-cell measurement of plasmid copy number and promoter activity
title_fullStr Single-cell measurement of plasmid copy number and promoter activity
title_full_unstemmed Single-cell measurement of plasmid copy number and promoter activity
title_short Single-cell measurement of plasmid copy number and promoter activity
title_sort single-cell measurement of plasmid copy number and promoter activity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7935883/
https://www.ncbi.nlm.nih.gov/pubmed/33674569
http://dx.doi.org/10.1038/s41467-021-21734-y
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