1 research outputs found
La restauración de la estación central sismológica de Tacubaya y su contexto
Techniques to analyze
human telomeres are imperative in studying
the molecular mechanism of aging and related diseases. Two important
aspects of telomeres are their length in DNA base pairs (bps) and
their biophysical nanometer dimensions. However, there are currently
no techniques that can simultaneously measure these quantities in
individual cell nuclei. Here, we develop and evaluate a telomere “dual”
gold nanoparticle-fluorescent probe simultaneously compatible with
both X-ray fluorescence (XRF) and super resolution microscopy. We
used silver enhancement to independently visualize the spatial locations
of gold nanoparticles inside the nuclei, comparing to a standard QFISH
(quantitative fluorescence <i>in situ</i> hybridization)
probe, and showed good specificity at ∼90%. For sensitivity,
we calculated telomere length based on a DNA/gold binding ratio using
XRF and compared to quantitative polymerase chain reaction (qPCR)
measurements. The sensitivity was low (∼10%), probably because
of steric interference prohibiting the relatively large 10 nm gold
nanoparticles access to DNA space. We then measured the biophysical
characteristics of individual telomeres using super resolution microscopy.
Telomeres that have an average length of ∼10 kbps, have diameters
ranging between ∼60–300 nm. Further, we treated cells
with a telomere-shortening drug and showed there was a small but significant
difference in telomere diameter in drug-treated vs control cells.
We discuss our results in relation to the current debate surrounding
telomere compaction