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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Springer Science and Business Media LLC
2021
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| Online Access: | https://hdl.handle.net/1721.1/133600 |
| _version_ | 1826206242456993792 |
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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 | MIT |
| 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. |
| first_indexed | 2024-09-23T13:26:21Z |
| format | Article |
| id | mit-1721.1/133600 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T13:26:21Z |
| publishDate | 2021 |
| publisher | Springer Science and Business Media LLC |
| record_format | dspace |
| spelling | mit-1721.1/1336002021-10-28T03:52:02Z Single-cell measurement of plasmid copy number and promoter activity Shao, Bin Rammohan, Jayan Anderson, Daniel A Alperovich, Nina Ross, David Voigt, Christopher A 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. 2021-10-27T19:53:45Z 2021-10-27T19:53:45Z 2021 2021-09-10T16:25:56Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/133600 en 10.1038/S41467-021-21734-Y Nature Communications Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf Springer Science and Business Media LLC Nature |
| spellingShingle | 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 |
| url | https://hdl.handle.net/1721.1/133600 |
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