Direct observation of frequency modulated transcription in single cells using light activation
Single-cell analysis has revealed that transcription is dynamic and stochastic, but tools are lacking that can determine the mechanism operating at a single gene. Here we utilize single-molecule observations of RNA in fixed and living cells to develop a single-cell model of steroid-receptor mediated...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2013-09-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/00750 |
| _version_ | 1811201091706028032 |
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| author | Daniel R Larson Christoph Fritzsch Liang Sun Xiuhau Meng David S Lawrence Robert H Singer |
| author_facet | Daniel R Larson Christoph Fritzsch Liang Sun Xiuhau Meng David S Lawrence Robert H Singer |
| author_sort | Daniel R Larson |
| collection | DOAJ |
| description | Single-cell analysis has revealed that transcription is dynamic and stochastic, but tools are lacking that can determine the mechanism operating at a single gene. Here we utilize single-molecule observations of RNA in fixed and living cells to develop a single-cell model of steroid-receptor mediated gene activation. We determine that steroids drive mRNA synthesis by frequency modulation of transcription. This digital behavior in single cells gives rise to the well-known analog dose response across the population. To test this model, we developed a light-activation technology to turn on a single steroid-responsive gene and follow dynamic synthesis of RNA from the activated locus. |
| first_indexed | 2024-04-12T02:16:03Z |
| format | Article |
| id | doaj.art-d00fedd288b642ef90927ea5b1ebe1fe |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:16:03Z |
| publishDate | 2013-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-d00fedd288b642ef90927ea5b1ebe1fe2022-12-22T03:52:16ZengeLife Sciences Publications LtdeLife2050-084X2013-09-01210.7554/eLife.00750Direct observation of frequency modulated transcription in single cells using light activationDaniel R Larson0Christoph Fritzsch1Liang Sun2Xiuhau Meng3David S Lawrence4Robert H Singer5Laboratory of Receptor Biology and Gene Expression, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, United StatesInstitute of Molecular Biology, Johannes Gutenberg University of Mainz, Mainz, GermanyDepartment of Chemistry, Division of Chemical Biology and Medicinal Chemistry, University of North Carolina, Chapel Hill, United States; Department of Pharmacology, University of North Carolina, Chapel Hill, United StatesDepartment of Anatomy and Structural Biology, Albert Einstein College of Medicine, New York, United StatesDepartment of Chemistry, Division of Chemical Biology and Medicinal Chemistry, University of North Carolina, Chapel Hill, United States; Department of Pharmacology, University of North Carolina, Chapel Hill, United StatesDepartment of Anatomy and Structural Biology, Albert Einstein College of Medicine, New York, United States; Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, New York, United StatesSingle-cell analysis has revealed that transcription is dynamic and stochastic, but tools are lacking that can determine the mechanism operating at a single gene. Here we utilize single-molecule observations of RNA in fixed and living cells to develop a single-cell model of steroid-receptor mediated gene activation. We determine that steroids drive mRNA synthesis by frequency modulation of transcription. This digital behavior in single cells gives rise to the well-known analog dose response across the population. To test this model, we developed a light-activation technology to turn on a single steroid-responsive gene and follow dynamic synthesis of RNA from the activated locus.https://elifesciences.org/articles/00750transcription factorgene expressionsingle-moleculefluorescence |
| spellingShingle | Daniel R Larson Christoph Fritzsch Liang Sun Xiuhau Meng David S Lawrence Robert H Singer Direct observation of frequency modulated transcription in single cells using light activation eLife transcription factor gene expression single-molecule fluorescence |
| title | Direct observation of frequency modulated transcription in single cells using light activation |
| title_full | Direct observation of frequency modulated transcription in single cells using light activation |
| title_fullStr | Direct observation of frequency modulated transcription in single cells using light activation |
| title_full_unstemmed | Direct observation of frequency modulated transcription in single cells using light activation |
| title_short | Direct observation of frequency modulated transcription in single cells using light activation |
| title_sort | direct observation of frequency modulated transcription in single cells using light activation |
| topic | transcription factor gene expression single-molecule fluorescence |
| url | https://elifesciences.org/articles/00750 |
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