ArticlePloS one2011
Single molecule in vivo analysis of toll-like receptor 9 and CpG DNA interaction.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
What it found
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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.
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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
16 citing papers in PubMed, 38 citations in OpenAlex.
- Article
- Control of A/D type CpG-ODN aggregates to a suitable size for induction of strong immunostimulant activity.Biochemistry and biophysics reports · 2023Article
- DNA-Based Biomaterials for Immunoengineering.Advanced healthcare materials · 2019Review
- Cell-free synthesis of human toll-like receptor 9 (TLR9): Optimization of synthesis conditions and functional analysis.PloS one · 2019Article
- Mitochondrial Damage-Associated Molecular Patterns of Injured Axons Induce Outgrowth of Schwann Cell Processes.Frontiers in cellular neuroscience · 2018Article
- Pharmacologic Protection of Mitochondrial DNA Integrity May Afford a New Strategy for Suppressing Lung Ischemia-Reperfusion Injury.Annals of the American Thoracic Society · 2017Review
- Article
- Epigenetic Toxicity of Trichloroethylene: A Single-Molecule Perspective.Toxicology research · 2016Article
- Dissecting the behavior and function of MBD3 in DNA methylation homeostasis by single-molecule spectroscopy and microscopy.Nucleic acids research · 2015Article
- Diversity of two forms of DNA methylation in the brain.Frontiers in genetics · 2014Article
- DNA-mediated assembly of protein heterodimers on membrane surfaces.Journal of the American Chemical Society · 2013Article
- Surface-enhanced Raman spectroscopy at single-molecule scale and its implications in biology.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2013Review
- Pattern recognition receptors in equine endotoxaemia and sepsis.Equine veterinary journal · 2012Article
- Single-molecule tools elucidate H2A.Z nucleosome composition.Journal of cell science · 2012Article
- DNA binding to proteolytically activated TLR9 is sequence-independent and enhanced by DNA curvature.The EMBO journal · 2012Article
- Inside single cells: quantitative analysis with advanced optics and nanomaterials.Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnologyReview
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 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Toll-like receptor 9 (TLR9) activates the innate immune system in response to oligonucleotides rich in CpG whereas DNA lacking CpG could inhibit its activation. However, the mechanism of how TLR9 interacts with nucleic acid and becomes activated in live cells is not well understood. Here, we report on the successful implementation of single molecule tools, constituting fluorescence correlation/cross-correlation spectroscopy (FCS and FCCS) and photon count histogram (PCH) with fluorescence lifetime imaging (FLIM) to study the interaction of TLR9-GFP with Cy5 labeled oligonucleotide containing CpG or lacking CpG in live HEK 293 cells. Our findings show that i) TLR9 predominantly forms homodimers (80%) before binding to a ligand and further addition of CpG or non CpG DNA does not necessarily increase the proportion of TLR9 dimers, ii) CpG DNA has a lower dissociation constant (62 nM±9 nM) compared to non CpG DNA (153 nM±26 nM) upon binding to TLR9, suggesting that a motif specific binding affinity of TLR9 could be an important factor in instituting a conformational change-dependant activation, and iii) both CpG and non CpG DNA binds to TLR9 with a 1∶2 stoichiometry in vivo. Collectively, through our findings we establish an in vivo model of TLR9 binding and activation by CpG DNA using single molecule fluorescence techniques for single cell studies.
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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.