ArticleMethods (San Diego, Calif.)2017
Quantifying transcription factor binding dynamics at the single-molecule level in live cells.
Article in Methods (San Diego, Calif.), 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 65 papers.
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Who cites it
65 citing papers in PubMed.
- Heparan Sulfate Controls Nanoscale Assembly of GPC3-Wnt Receptor Complexes.bioRxiv : the preprint server for biology · 2026Article
- QuantiTrack: A unified software to study protein dynamics in living cells.bioRxiv : the preprint server for biology · 2026Article
- Article
- Single-Molecule Localization Microscopy in the Nucleus with Self-Labelling Tags: Halo, SNAP, and CLIP Tags.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Synthetic fluorophores for live-cell fluorescence microscopy and biosensing.Nature chemical biology · 2025Review
- Single-cell synaptome mapping of endogenous protein subpopulations in mammalian brain.Nature communications · 2025Article
- Thioether editing generally increases the photostability of rhodamine dyes on self-labeling tags.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Transcription factors form a ternary complex with NIPBL/MAU2 to localize cohesin at enhancers.Nucleic acids research · 2025Article
- Zooming into Gene Activation: Estrogen Receptor α Dimerization and DNA Binding Visualized by High-Speed Atomic Force Microscopy.ACS nano · 2025Article
- Experimental Approaches to Visualize Effector Protein Translocation During Host-Pathogen Interactions.BioEssays : news and reviews in molecular, cellular and developmental biology · 2025Review
- Single-molecule tracking in living microbial cells.Biophysics reports · 2025Article
- Transcription dynamics and genome organization in the mammalian nucleus: Recent advances.Molecular cell · 2025Review
- A high-throughput platform for single-molecule tracking identifies drug interaction and cellular mechanisms.eLife · 2025Article
- Pre-ribosomal particles from nucleoli to cytoplasm.Nucleus (Austin, Tex.) · 2024Review
- Real-time imaging of axonal membrane protein life cycles.Nature protocols · 2024Review
- Chromatin structure and dynamics: one nucleosome at a time.Histochemistry and cell biology · 2024Review
- Article
- BenzoHTag, a fluorogenic self-labeling protein developed using molecular evolution.bioRxiv : the preprint server for biology · 2024Article
- Analyzing paramagnetic NMR data on target DNA search by proteins using a discrete-state kinetic model for translocation.Biopolymers · 2024Article
- Article
5 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors.
Funding
Abstract
Progressive, technological achievements in the quantitative fluorescence microscopy field are allowing researches from many different areas to start unraveling the dynamic intricacies of biological processes inside living cells. From super-resolution microscopy techniques to tracking of individual proteins, fluorescence microscopy is changing our perspective on how the cell works. Fortunately, a growing number of research groups are exploring single-molecule studies in living cells. However, no clear consensus exists on several key aspects of the technique such as image acquisition conditions, or analysis of the obtained data. Here, we describe a detailed approach to perform single-molecule tracking (SMT) of transcription factors in living cells to obtain key binding characteristics, namely their residence time and bound fractions. We discuss different types of fluorophores, labeling density, microscope, cameras, data acquisition, and data analysis. Using the glucocorticoid receptor as a model transcription factor, we compared alternate tags (GFP, mEOS, HaloTag, SNAP-tag, CLIP-tag) for potential multicolor applications. We also examine different methods to extract the dissociation rates and compare them with simulated data. Finally, we discuss several challenges that this exciting technique still faces.
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Registered trials
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