Ultrastructure and nuclear architecture of telomeric chromatin revealed by correlative light and electron microscopy.

Telomeres, the ends of linear chromosomes, are composed of repetitive DNA sequences, histones and a protein complex called shelterin. How DNA is packaged at telomeres is an outstanding question in the field with significant implications for human health and disease. Here, we studied the architecture of telomeres and their spatial association with other chromatin domains in different cell types using correlative light and electron microscopy. To this end, the shelterin protein TRF1 or TRF2 was fused in tandem to eGFP and the peroxidase APEX2, which provided a selective and electron-dense label to interrogate telomere organization by transmission electron microscopy, electron tomography and scanning electron microscopy. Together, our work reveals, for the first time, ultrastructural insight into telomere architecture. We show that telomeres are composed of a dense and highly compacted mesh of chromatin fibres. In addition, we identify marked differences in telomere size, shape and chromatin compaction between cancer and non-cancer cells and show that telomeres are in direct contact with other heterochromatin regions. Our work resolves the internal architecture of telomeres with unprecedented resolution and advances our understanding of how telomeres are organized in situ.

Upconversion Nanoparticle Powered Microneedle Patches for Transdermal Delivery of siRNA.

Microneedles (MNs) permit the delivery of nucleic acids like small interfering RNA (siRNA) through the stratum corneum and subsequently into the skin tissue. However, skin penetration is only the first step in successful implementation of siRNA therapy. These delivered siRNAs need to be resistant to enzymatic degradation, enter target cells, and escape the endosome-lysosome degradation axis. To address this challenge, this article introduces a nanoparticle-embedding MN system that contains a dissolvable hyaluronic acid (HA) matrix and mesoporous silica-coated upconversion nanoparticles (UCNPs@mSiO2 ). The mesoporous silica (mSiO2 ) shell is used to load and protect siRNA while the upconversion nanoparticle (UCNP) core allows the tracking of MN skin penetration and NP diffusion through upconversion luminescence imaging or optical coherence tomography (OCT) imaging. Once inserted into the skin, the HA matrix dissolves and UCNPs@mSiO2 diffuse in the skin tissue before entering the cells for delivering the loaded genes. As a proof of concept, this system is used to deliver molecular beacons (MBs) and siRNA targeting transforming growth factor-beta type I receptor (TGF-ßRI) that is potentially used for abnormal scar treatment.