Carolin Leonhardt

German

Non-viral gene delivery is of growing interest for gene therapy applications and basic research. My PhD project aims to quantitatively study the delivery of plasmid DNA (pDNA), messenger RNA (mRNA) and small interfering RNA (siRNA) to cells. Using time-lapse fluorescence microscopy and fluorescent reporter proteins, the intensity levels of numerous cells are monitored after transfection with pDNA/mRNA/siRNA. In order to allow for high-throughput, automated image analysis, I use micro-patterning techniques to structure cell growth substrates with islands of cell-adhesive proteins. Gene expression levels, the timing of expression onset/knockdown and kinetic rates are analyzed using image analysis techniques as well as kinetic rate models to gain insights into the processes underlying artificial gene-transfer. All observed parameters are analyzed quantitatively at the single-cell level. This single-cell approach reveals the stochastic behaviour of cells that would be obscured in ensemble measurements.