Sukanya Punthambaker
Wyss Institute for Biologically Inspired Engineering at Harvard University, USA
Title: Nanopore sequencing for next generation personalized genomic medicine
Biography
Biography: Sukanya Punthambaker
Abstract
Nanotechnology is at the fore front of precision medicine for diagnostics and targeted medicine. Sequencing the genome allows the understanding of the genetic make-up of an individual and to predict the susceptibility to a disease thus paving the way for therapeutics and personalized medicine. Nanopore technology is a fourth generation sequencing technology that offers a single molecule sequencing platform which can be highly scalable for high through put DNA sequencing. This technology is a powerful tool that can revolutionize personalized genomic medicine since it’s fast, cost-effective, requires less amount of sample material and no amplification steps. We recently reported a nanopore-based sequencing by synthesis technology wherein we show that a polymerase could be coupled to the α-hemolysin pore embedded in a lipid bilayer formed on an electrode array. Using this method, we demonstrate that DNA sequencing could be achieved at a single molecule level using tagged nucleotides that get captured in the pore yielding signature current blockade events. Here, we use this nanopore platform to describe a method to rapidly screen polymerase variants in a multiplex and high-throughput manner. We utilize barcoded DNA templates to uniquely identify DNA polymerase variants in a heterogeneous population. We have also developed a robust classification algorithm that discriminates kinetic characteristics of the different polymerase mutants. Our method can be broadly applicable to a variety of enzyme screening techniques