Dresden 2006 – scientific programme

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AKB: Biologische Physik

AKB 21: Intracellular Transport

AKB 21.6: Talk

Thursday, March 30, 2006, 12:00–12:15, ZEU 260

Fluorescence Microscopy reveals the mechanistic details of nano-sized gene carrier transport in living cells — •Ralf Bausinger¹, Nadia Ruthardt¹, Karla de Bruin¹, Katharina von Gersdorff², Manfred Ogris², Ernst Wagner², Andreas Zumbusch¹, and Christoph Bräuchle¹ — ¹Department of Chemistry and Biochemistry, LMU München, Butenandtstr. 5-13, 81377 München — ²Department of Pharmacy, LMU München, Butenandtstr. 5-13, 81377 München

Non-viral vectors consisting of the cationic polymer polyethyleneimine (PEI) and plasmid DNA are widely used for gene delivery into living cells. A detailed knowledge about the different stages which occur during the polyplex entry into the cell and its nucleus are prerequisite for further optimising the transfection process. We use highly sensitive fluorescence wide-field microscopy techniques to visualise the interaction of PEI/DNA polyplexes with the eGFP-labeled actin and tubulin cytoskeleton of living Huh-7 cells. Besides normal diffusion within the cell membrane we observe anomalous diffusion of the polyplexes due to their interaction with actin filaments as well as active directed transport along the microtubules [1]. In long-term experiments during mitosis we investigate the association of polyplexes to the spindle apparatus as a possible nuclear entry mechanism. We also compare the behaviour of these classical PEI/DNA polyplexes to more advanced non-viral vectors with polyethyleneglycol shielding and epidermal growth factor targeting.

[1] Bausinger et al., Angew. Chem., accepted