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Time-resolved analysis of transcription kinetics in single live mammalian cells
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Choi, Hongyoung | - |
dc.contributor.author | Lee, Byung Hun | - |
dc.contributor.author | Park, Hye Yoon | - |
dc.date.accessioned | 2023-01-02T02:36:07Z | - |
dc.date.available | 2023-01-02T02:36:07Z | - |
dc.date.created | 2022-12-02 | - |
dc.date.issued | 2022-11 | - |
dc.identifier.citation | Frontiers in Physics, Vol.10, p. 977125 | - |
dc.identifier.issn | 2296-424X | - |
dc.identifier.uri | https://hdl.handle.net/10371/188764 | - |
dc.description.abstract | In eukaryotic cells, RNA polymerase II synthesizes mRNA in three stages, initiation, elongation, and termination, and numerous factors determine how quickly a gene is transcribed to produce mRNA molecules through these steps. However, there are few techniques available to measure the rate of each step in living cells, which prevents a better understanding of transcriptional regulation. Here, we present a quantitative analysis method to extract kinetic rates of transcription from time-lapse imaging data of fluorescently labeled mRNA in live cells. Using embryonic fibroblasts cultured from two knock-in mouse models, we monitored transcription of beta-actin and Arc mRNA labeled with MS2 and PP7 stem-loop systems, respectively. After inhibiting transcription initiation, we measured the elongation rate and the termination time by fitting the time trace of transcription intensity with a mathematical model function. We validated our results by comparing them with those from an autocorrelation analysis and stochastic simulations. This live-cell transcription analysis method will be useful for studying the regulation of elongation and termination steps, providing insight into the diverse mechanisms of transcriptional processes. | - |
dc.language | 영어 | - |
dc.publisher | Frontiers Media S.A. | - |
dc.title | Time-resolved analysis of transcription kinetics in single live mammalian cells | - |
dc.type | Article | - |
dc.identifier.doi | 10.3389/fphy.2022.977125 | - |
dc.citation.journaltitle | Frontiers in Physics | - |
dc.identifier.wosid | 000885271400001 | - |
dc.identifier.scopusid | 2-s2.0-85142089606 | - |
dc.citation.startpage | 977125 | - |
dc.citation.volume | 10 | - |
dc.description.isOpenAccess | Y | - |
dc.contributor.affiliatedAuthor | Park, Hye Yoon | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
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