ArticleMethods in molecular biology (Clifton, N.J.)2013
Measuring transcription dynamics in living cells using fluctuation analysis.
Article in Methods in molecular biology (Clifton, N.J.), 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 37 citations in OpenAlex.
- Imaging Retroviral RNA Genome Heterodimers Using Bimolecular Fluorescence Complementation (BiFC).Viruses · 2025Article
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- Unified bursting strategies in ectopic and endogenousbioRxiv : the preprint server for biology · 2024Article
- BurstDECONV: a signal deconvolution method to uncover mechanisms of transcriptional bursting in live cells.Nucleic acids research · 2023Article
- Long-term imaging of individual mRNA molecules in living cells.Cell reports methods · 2022Article
- Nuclear envelope mechanobiology: linking the nuclear structure and function.Nucleus (Austin, Tex.) · 2021Review
- Quantitative imaging of transcription in living Drosophila embryos reveals the impact of core promoter motifs on promoter state dynamics.Nature communications · 2021Article
- Enhanced detection of expanded repeat mRNA foci with hybridization chain reaction.Acta neuropathologica communications · 2021Article
- Unraveling the molecular interactions involved in phase separation of glucocorticoid receptor.BMC biology · 2020Article
- Mammalian gene expression variability is explained by underlying cell state.Molecular systems biology · 2020Article
- Article
- Single-gene dual-color reporter cell line to analyze RNA synthesis in vivo.Methods (San Diego, Calif.) · 2016Article
- Fluctuation Analysis: Dissecting Transcriptional Kinetics with Signal Theory.Methods in enzymology · 2016Article
- Kinetic competition during the transcription cycle results in stochastic RNA processing.eLife · 2014Article
Corrections and comments
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
Abstract
Single-cell studies of gene regulation suggest that transcription dynamics play a fundamental role in determining expression heterogeneity within a population. In addition, the three-dimensional organization of the nucleus seems to both reflect and influence expression patterns in the cell. Therefore, to gain a holistic understanding of transcriptional regulation, it is necessary to develop methods for studying transcription of single genes in living cells with high spatial and temporal resolution. In this chapter, we describe a recently developed approach for visualizing and quantifying pre-mRNA synthesis at a single active gene in the nucleus. The approach is based on the high-affinity interaction between MS2/PP7 bacteriophage coat proteins and RNA hairpins which are transcribed by the gene of interest. The MS2/PP7 coat protein is fused to a fluorescent protein and binds the nascent mRNA, allowing for detection of single transcription events in the fluorescence microscope. By time-lapse fluorescence imaging and quantitative image analysis, one can generate a time trace of fluorescence intensity at the site of transcription. By temporal autocorrelation analysis, one can determine enzymatic activities of RNAP such as initiation rate and elongation rate. In this protocol, we summarize the experimental concept, design, and execution for real-time observation of transcription in living cells.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.