Publications by authors named "Dave Klenerman"
A major challenge in single-molecule imaging is tracking the dynamics of proteins or complexes for long periods of time in the dense environments found in living cells. Here, we introduce the concept of using FRET to enhance the photophysical properties of photo-modulatable (PM) fluorophores commonly used in such studies. By developing novel single-molecule FRET pairs, consisting of a PM donor fluorophore (either mEos3.
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- Researchers developed a new protocol that combines fluorescence imaging and Hi-C assays to study genome organization in individual cells.
- This method allows for the simultaneous capture of chromosome contacts and fluorescence images, providing comprehensive data on genome structures with high resolution.
- The entire process of preparing single-cell Hi-C libraries takes about 5 days for experienced researchers, requiring basic knowledge of microscopy and bioinformatics for data analysis.
Article Synopsis
- The folding of genomic DNA is essential for nuclear functions, and researchers have developed a new technique to visualize and study individual mammalian genomes in 3D.
- This method allows scientists to observe genome folding at a scale smaller than 100kb, revealing that individual structures vary between cells while certain genomic features have consistent organization across cells.
- By examining gene regulation mechanisms, this research demonstrates how single-cell genomic structure can provide insights into biological processes.
HIV-1 genomic RNA has a noncoding 5' region containing sequential conserved structural motifs that control many parts of the life cycle. Very limited data exist on their three-dimensional (3D) conformation and, hence, how they work structurally. To assemble a working model, we experimentally reassessed secondary structure elements of a 240-nt region and used single-molecule distances, derived from fluorescence resonance energy transfer, between defined locations in these elements as restraints to drive folding of the secondary structure into a 3D model with an estimated resolution below 10 Å.
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