ArticleScientific reports2016
Super-resolution imaging of fluorescently labeled, endogenous RNA Polymerase II in living cells with CRISPR/Cas9-mediated gene editing.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers.
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Who cites it
41 citing papers in PubMed.
- Resolving Sub-Nuclear Architecture from Compartments to Functional Domains.International journal of molecular sciences · 2026Review
- RNA polymerase II transcription compartments - from factories to condensates.Nature reviews. Genetics · 2025Review
- Transcriptional Condensates: Epigenetic Reprogramming and Therapeutic Targets in Hematologic Malignancies.Cancers · 2025Review
- Article
- Illuminating biomolecular assemblies in gene regulation.Biophysical reviews · 2025Review
- The role of transcription bodies in gene expression: what embryos teach us.Biochemical Society transactions · 2025Review
- Article
- ULK1 forms distinct oligomeric states and nanoscopic structures during autophagy initiation.Science advances · 2023Article
- Combined SPT and FCS methods reveal a mechanism of RNAP II oversampling in cell nuclei.Scientific reports · 2023Article
- Review
- Autophagy receptor NDP52 alters DNA conformation to modulate RNA polymerase II transcription.Nature communications · 2023Article
- Article
- Spatially coherent diffusion of human RNA Pol II depends on transcriptional state rather than chromatin motion.Nucleus (Austin, Tex.) · 2022Article
- Superresolution microscopy localizes endogenous Dvl2 to Wnt signaling-responsive biomolecular condensates.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Single-molecule tracking (SMT) and localization of SRF and MRTF transcription factors during neuronal stimulation and differentiation.Open biology · 2022Article
- Myosin VI regulates the spatial organisation of mammalian transcription initiation.Nature communications · 2022Article
- MYC amplifies gene expression through global changes in transcription factor dynamics.Cell reports · 2022Article
- CTCF-mediated chromatin looping provides a topological framework for the formation of phase-separated transcriptional condensates.Nucleic acids research · 2022Article
- Towards a 'Spot On' Understanding of Transcription in the Nucleus.Journal of molecular biology · 2021Review
- How Single-Molecule Localization Microscopy Expanded Our Mechanistic Understanding of RNA Polymerase II Transcription.International journal of molecular sciences · 2021Review
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6 authors.
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Abstract
Live cell imaging of mammalian RNA polymerase II (Pol II) has previously relied on random insertions of exogenous, mutant Pol II coupled with the degradation of endogenous Pol II using a toxin, α-amanitin. Therefore, it has been unclear whether over-expression of labeled Pol II under an exogenous promoter may have played a role in reported Pol II dynamics in vivo. Here we label the endogenous Pol II in mouse embryonic fibroblast (MEF) cells using the CRISPR/Cas9 gene editing system. Using single-molecule based super-resolution imaging in the living cells, we captured endogenous Pol II clusters. Consistent with previous studies, we observed that Pol II clusters were short-lived (cluster lifetime ~8 s) in living cells. Moreover, dynamic responses to serum-stimulation, and drug-mediated transcription inhibition were all in agreement with previous observations in the exogenous Pol II MEF cell line. Our findings suggest that previous exogenously tagged Pol II faithfully recapitulated the endogenous polymerase clustering dynamics in living cells, and our approach may in principle be used to directly label transcription factors for live cell imaging.
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