ArticleBioconjugate chemistry2011
Hairpin-like fluorescent probe for imaging of NF-κB transcription factor activity.
Article in Bioconjugate chemistry, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 13 citations in OpenAlex.
- Imaging NF-κB activity in a murine model of early stage diabetes.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2020Article
- Self-neutralizing oligonucleotides with enhanced cellular uptake.Organic & biomolecular chemistry · 2017Article
- Fluorocarbons Enhance Intracellular Delivery of Short STAT3-sensors and Enable Specific Imaging.Theranostics · 2017Article
- The three-dimensional context of a double helix determines the fluorescence of the internucleoside-tethered pair of fluorophores.Molecular bioSystems · 2013Article
- DNA-based sensor for real-time measurement of the enzymatic activity of human topoisomerase I.Sensors (Basel, Switzerland) · 2013Article
- Sensing of transcription factor binding via cyanine dye pair fluorescence lifetime changes.Molecular bioSystems · 2012Article
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Authors and funding
4 authors at 3 institutions in 2 countries.
Funding
Abstract
Three oligodeoxyribonucleotides (ODN) covalently labeled with near-infrared (NIR) fluorochromes were synthesized and characterized with a goal of comparing in vitro a hairpin-based and a duplex-based FRET probe designed for the detection of human recombinant NF-κB p50/p65 heterodimer binding to DNA. Using deoxyguanosine phosphoramidite with a phosphorus-linked aminoethylene (diethylene glycol) hydrophilic linker, we synthesized ODNs with internucleoside reactive sites. The hairpin loop amino linker was modified with IRDye 800CW (FRET acceptor), and the 3'-end was modified with Cy5.5 (FRET donor) using a dithio-linker. To obtain a duplex probe, we conjugated Cy5.5 and 800CW to complementary strands at the distance of ten base pairs in the resultant duplex. No quenching of dyes was observed in either probe. The FRET efficiency was higher in the duplex (71%) than in the hairpin (56%) due to a more favorable distance between the donor and the acceptor. However, the hairpin design allowed more precise ratiometric measurement of fluorescence intensity changes as a result of NF-κB p50/p65 binding to the probe. We determined that as a result of binding there was a statistically significant increase of fluorescence intensity of Cy5.5 (donor) due to a decrease of FRET if normalized by 800CW intensity measured independently of FRET. We conclude that the hairpin based probe design allows for the synthesis of a dual fluorescence imaging probe that renders signal changes that are simple to interpret and stoichiometrically correct for detecting transcription factor-DNA interactions.
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Registered trials
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