Characterisation of DNA delivery through fluctuation correlation spectroscopy

Abstract

Gene delivery or gene therapy involves the coupling of a nucleic acid sequences with a delivery vehicle, such as viruses or lipids, to facilitate uptake into the target cell. During the delivery of the exogenous nucleic acid sequences a number of obstacles are faced. As a result there has been a significant emphasis on developing an understanding of the processes involved. To date most studies have focused on limited time intervals or small regions of the cell, or fixed samples. Developing a realtime, multi-level information approach to characterise the live cell responses to exogenous nucleic acid sequences is expected to underpin optimising Gene Therapy methodology. In addition, it is likely this approach could be applied to drug delivery and enhance our understanding of cellular processing. Fluorescence Correlation Spectroscopy (FCS) is one approach to analyse single molecules of extremely low concentration with high spatial and temporal resolution (for example, 600 particles are expected to be within the illumination volume at a given time in a 1 microM solution of a small molecule). Compared to other fluorescence-based approaches FCS measures the spontaneous intensity fluctuations caused by minute changes in particle behaviour, rather than relying on the emission intensity of fluorescence. Fundamentally, FCS has the potential to measure and fully quantify all physical processes occurring in a cell that give rise to fluctuations in the fluorescence signal including mobility and aggregation.

Date of Award	2015
Original language	English