ArticleNature protocols2013
Quantifying the transcriptional output of single alleles in single living mammalian cells.
Article in Nature protocols, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
20 citing papers in PubMed, 33 citations in OpenAlex.
- Measuring the Proximity of Nuclear lncRNAs to Nuclear Bodies.Methods in molecular biology (Clifton, N.J.) · 2027Article
- Insights into RNA-mediated pathology in new mouse models of Huntington's disease.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2024Article
- A rapid inducible RNA decay system reveals fast mRNA decay in P-bodies.Nature communications · 2024Article
- Light chain 2 is a Tctex-type related axonemal dynein light chain that regulates directional ciliary motility in Trypanosoma brucei.PLoS pathogens · 2022Article
- Gene transcription in bursting: a unified mode for realizing accuracy and stochasticity.Biological reviews of the Cambridge Philosophical Society · 2019Article
- S-phase transcriptional buffering quantified on two different promoters.Life science alliance · 2018Article
- Dynamic imaging demonstrates that pulsed electromagnetic fields (PEMF) suppress IL-6 transcription in bovine nucleus pulposus cells.Journal of orthopaedic research : official publication of the Orthopaedic Research Society · 2018Article
- CD-tagging-MS2: detecting allelic expression of endogenous mRNAs and their protein products in single cells.Biology methods & protocols · 2017Article
- Article
- DTIE, a novel core promoter element that directs start site selection in TATA-less genes.Nucleic acids research · 2016Article
- Random monoallelic expression of autosomal genes: stochastic transcription and allele-level regulation.Nature reviews. Genetics · 2015Review
- Cellular Levels of Signaling Factors Are Sensed by β-actin Alleles to Modulate Transcriptional Pulse Intensity.Cell reports · 2015Article
- Design (and) principles of nuclear dynamics in Stockholm.Nucleus (Austin, Tex.) · 2015Article
- Stochastic promoter activation affects Nanog expression variability in mouse embryonic stem cells.Scientific reports · 2014Article
- High-throughput microfluidic single-cell analysis pipeline for studies of signaling dynamics.Nature protocols · 2014Article
- Subcellular mRNA localisation at a glance.Journal of cell science · 2014Review
- Seeing is believing: visualizing transcriptional dynamics in single cells.Journal of cellular physiology · 2014Review
- Advanced fluorescence microscopy methods for the real-time study of transcription and chromatin dynamics.Transcription · 2014Review
- RNA imaging in living cells - methods and applications.RNA biology · 2014Review
- Zooming in on single active genes in living mammalian cells.Histochemistry and cell biology · 2013Review
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Transcription kinetics of actively transcribing genes in vivo have generally been measured using tandem gene arrays. However, tandem arrays do not reflect the endogenous state of genome organization in which genes appear as single alleles. Here we present a robust technique for the quantification of mRNA synthesis from a single allele in real time in single living mammalian cells. The protocol describes how to generate cell clones harboring an MS2-tagged allele and how to detect in vivo transcription from this tagged allele at high spatial and temporal resolution throughout the cell cycle. Quantification of nascent mRNAs produced from the single tagged allele is performed using RNA fluorescence in situ hybridization (FISH) and live-cell imaging. Subsequent analyses and data modeling detailed in the protocol include measurements of transcription rates of RNA polymerase II, determination of the number of polymerases recruited to the tagged allele and measurement of the spacing between polymerases. Generation of the cells containing the single tagged alleles should take up to 1 month; RNA FISH or live-cell imaging will require an additional week.
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Registered trials
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.