Dresden 2006 – wissenschaftliches Programm

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AKB: Biologische Physik

AKB 30: Poster Session I

AKB 30.26: Poster

Montag, 27. März 2006, 15:30–18:00, P1

Targeted transfection and gene-silencing using femtosecond laser pulses — •Elke Haustein, Thomas Ohrt, and Petra Schwille — TU Dresden, Institute for Biophysics, Tatzberg 47-51, D-01307 Dresden

To manipulate cellular function and morphology, molecules that can specifically affect intracellular processes have to be transported through the plasma membrane. Small interfering RNAs are key intermediates of a post-transcriptional gene-silencing mechanism known as RNA interference (RNAi). This technique allows for a temporary, easy-to-use and specific protein knockdown and thus can be applied both to biological research and future therapeutic applications. To achieve optimal results, controlling both the kinetics of RNA delivery and the final amount of probe substance within the target cell is mandatory. So far, different techniques have been tested to deliver siRNAs in situ, most of which depend on cellular uptake mechanisms. But to avoid delivery-related artefacts, the siRNAs have to enter the cytoplasm directly. Therefore, we propose an alternative means to deliver RNA to cells. Using a confocal setup, the parallel infrared laser beam is directed into an inverted microscope and focussed onto the cell membrane. The cells are then exposed to femtosecond laser pulses for varying time intervals. The resulting perforation of the plasma membrane allows uptake of the RNA added to the surrounding medium. Using a fluorescence-based assay, the effectivity of this No-dqphotoporationNo-dq approach can be characterised by fluorescence-based techniques as well as with biochemical means.