Publications by authors named "V Buckle"

Eukaryotic genomes are organized by loop extrusion and sister chromatid cohesion, both mediated by the multimeric cohesin protein complex. Understanding how cohesin holds sister DNAs together, and how loss of cohesion causes age-related infertility in females, requires knowledge as to cohesin's stoichiometry in vivo. Using quantitative super-resolution imaging, we identified two discrete populations of chromatin-bound cohesin in postreplicative human cells.

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Determining the mechanisms by which genes are switched on and off during development is a key aim of current biomedical research. Gene transcription has been widely observed to occur in a discontinuous fashion, with short bursts of activity interspersed with periods of inactivity. It is currently not known if or how this dynamic behaviour changes as mammalian cells differentiate.

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DNA fluorescence in situ hybridization (FISH) has been a central technique in advancing our understanding of how chromatin is organized within the nucleus. With the increasing resolution offered by super-resolution microscopy, the optimal maintenance of chromatin structure within the nucleus is essential for accuracy in measurements and interpretation of data. However, standard 3D-FISH requires potentially destructive heat denaturation in the presence of chaotropic agents such as formamide to allow access to the DNA strands for labeled FISH probes.

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Article Synopsis
  • Chromosome conformation capture (3C) is a versatile tool for exploring biological questions, but existing methods either offer low-resolution data for the entire genome or higher resolution at a few specific locations.
  • This study presents two key improvements to 3C techniques, significantly reducing noise from reporter events by isolating nuclei after ligation.
  • Using Nuclear-Titrated Capture-C, the authors achieved reproducible, high-resolution genome-wide interaction profiles, focusing on 8055 gene promoters in erythroid cells, and examined the influence of promoter hubs and super-enhancers on gene regulation.
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