Techniques

We use several molecular biology techniques including restriction-based molecular cloning, PCR, site-directed mutagenesis, DNA/RNA isolation from human cells, cDNA synthesis, real-time RT-qPCR etc…

We express proteins/complexes recombinantly in various systems including E. coli, baculovirus-infected insect cells, and cultured human cells for use in biochemical and X-ray crystallographic experiments.

We employ a wide range of biochemical assays for determining binding constants, enzymatic activity etc. of various telomeric and telomerase complexes. Some examples are shown below.

We host a state-of-the-art Size-exclusion Chromatography coupled to Multi-angle Light Scattering instruments (from Wyatt, FPLC from Cytiva/GE) for determining the molecular weight and stoichiometry of native proteins, protein-protein complexes, and protein-nucleic acid complexes

We use X-ray crystallography as a major tool for determining the molecular bases of the various biological and biochemical phenomena that we investigate. We are embarking on negative-stain EM and cryo-EM to further expand our expertise in structural biology.

We use a wide range of cell biological assays ranging from immunofluorescence (IF) and fluorescence in situ hybridization (FISH) imaging of fixed human cells in culture, telomere length analysis, immunoprecipitation (IP), ChIP etc… We also use state-of-the-art techniques to engineer cell lines expressing genetic constructs of our interests in a stable and controlled fashion. Most recently we have successfully used CRISPR-Cas9 technology to introduce both knock out and knock in (single-site changes) mutations in telomeric genes in cultured human cells. Some examples of methods used in the lab are shown below.

We have successfully optimized both knock-out and knock-in of genes in cultured cells (like HEK 293T and HeLa) using CRISPR-Cas9 technology. Knock-in changes include point mutations as well as the insertion of epitope tags such as FLAG tags.