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SYBP: Biophotonik

SYBP 3: Einzelmolekülmethoden III

SYBP 3.2: Vortrag

Donnerstag, 7. März 2002, 17:00–17:15, HS 11/215

Single-Virus-Tracing: Infection Pathway of AAV — •Stephan Grimm¹, Thomas Endress¹, Georg Seisenberger¹, Hildegard Büning², Martin Ried², Michael Hallek² und Christoph Bräuchle¹ — ¹Department Chemie, Lehrstuhl Physikalische Chemie I und Center for Nanoscience, Ludwig-Maximilians-Universität München, Butenandtstr. 5 - 13, 81377 München — ²Genzentrum der Universität München, Feodor-Lynen-Str. 25

Single molecule detection is a novel technique for the investigation of biological processes on the molecular level. The study of individual molecules allows one to look beyond ensemble-averaged properties, thus revealing details of the molecular behavior which are otherwise hidden by spatial, spectral and temporal averaging in ensemble measurements. Here we show that this method can be used for the kinetic characterization of the infection pathway of a single dye-labeled virus. Single molecule experiments were conducted to follow the migration of individual Adeno-associated viruses on their infectious entry pathway into a living HeLa cell. Only one fluorescent dye molecule (Cy5 or Cy3 respectively) was attached to a virus in order not to influence its physiological behavior. The fluorescent dye-label allows one to observe the movement of a virus. Newest results forming a detailed picture of the processes involved in the uptake of a virus in a living cell could be observed. This is of relevance to an understanding of the pathogenic mechanisms of a virus infection. In addition single virus tracing experiments can be applied to both improvement of limitations of virus-mediated gene delivery in living cells (e.g. gene therapy) and development of antiviral drugs.