Evidence map›Paper›PMID 26914315›Full record

ArticleNature protocols2016

Isolation of the protein and RNA content of active sites of transcription from mammalian cells.

Svitlana Melnik, Maïwen Caudron-Herger, Lilija Brant, Ian M Carr, Karsten Rippe, Peter R Cook, Argyris Papantonis

Open access · greenAbstract read
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In one paragraph

Article in Nature protocols, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.

0numbers the graph read from it
0cells of the map it votes in
9citing papers in PubMed
1.7field-weighted citation impact, top 16% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

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.

2 · The registry

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.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

9 citing papers in PubMed, 24 citations in OpenAlex.

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  7. MiR-181a TargetsFrontiers in cell and developmental biology · 2021
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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors at 4 institutions in 2 countries.

Svitlana MelnikSir William Dunn School of Pathology, University of Oxford, Oxford, UK.
Maïwen Caudron-HergerGerman Cancer Research Center (DKFZ) and BioQuant, Heidelberg, Germany.
Lilija BrantCenter for Molecular Medicine, University of Cologne, Cologne, Germany.
Ian M CarrInstitute of Biomedical and Clinical Sciences, University of Leeds, Leeds, UK.
Karsten RippeGerman Cancer Research Center (DKFZ) and BioQuant, Heidelberg, Germany.ORCID http://orcid.org/0000-0001-9951-9395
Peter R CookSir William Dunn School of Pathology, University of Oxford, Oxford, UK.
Argyris PapantonisCenter for Molecular Medicine, University of Cologne, Cologne, Germany.
German Cancer Research Center · DEUniversity of Cologne · DEUniversity of Oxford · GBUniversity of Leeds · GB

Funding

Medical Research Council
6 · The paper itself

Abstract

Mammalian cell nuclei contain three RNA polymerases (RNAP I, RNAP II and RNAP III), which transcribe different gene subsets, and whose active forms are contained in supramolecular complexes known as 'transcription factories.' These complexes are difficult to isolate because they are embedded in the 3D structure of the nucleus. Factories exchange components with the soluble nucleoplasmic pool over time as gene expression programs change during development or disease. Analysis of their content can provide information on the nascent transcriptome and its regulators. Here we describe a protocol for the isolation of large factory fragments under isotonic salt concentrations in <72 h. It relies on DNase I-mediated detachment of chromatin from the nuclear substructure of freshly isolated, unfixed cells, followed by caspase treatment to release multi-megadalton factory complexes. These complexes retain transcriptional activity, and isolation of their contents is compatible with downstream analyses by mass spectrometry (MS) or RNA-sequencing (RNA-seq) to catalog the proteins and RNA associated with sites of active transcription.

Indexed as

Gene Expression ProfilingTranscription, GeneticAnimalsCell FractionationCell LineCell NucleusCHO CellsCricetulusHeLa CellsHumansHuman Umbilical Vein Endothelial CellsMass SpectrometryProteinsProteomicsRNASequence Analysis, RNAProteinsRNA

Identifiers

PMID26914315
OpenAlexW2275420586

What OpenQuestion holds

Textmetadata
Read underepoch 390

Registered trials

None linked

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.