Supplemental information

PhD thesis: “Visualization of the intracellular trafficking of Adeno-associated virus type 2”

Kerstin Lux

 

1. Movies:

Movie 1 chapter 2.1.4: HeLa dsRed Nuc cells infected with 100%-GFP-VP2-AAV (approx. 106 capsids/cell). 

Movie generated by conventional time laps microscopy:

27 images were taken with a time laps of 30 seconds.

Some GFP-tagged AAV virions seem to touch the cell membrane multiples times. Most virions stack to the membrane, but some GFP tagged virions seem to move inside the cytoplasm. These observations confirm previous observations made by SVT.

      link\movie 1.AVI

 

Movies 2-4 chapter 2.1.10

Movies 2 and 3: HeLa cells infected with Cy5 labeled AAV (approx. 300 capsids/cell).

Real time movies generated by SVT measurements: directed movements of Cy5 labeled AAV inside HeLa cells.

Images were taken every 40 milliseconds. Movie 2 contains 200 and movie 3 contains 110 frames.

link\movie 2.AVI

link\movie 3.AVI

 

Movie 4: PtK2 cells infected with 100%-GFP-VP2-AAV (approx. 106 capsids/cell).

Movie generated by conventional time laps microscopy:

Images were taken with a time laps of 2 seconds, 100 images were taken.

GFP tagged AAV particles can be observed performing directed movements. Furthermore, many particles perform small, oscillatory movements without making large displacements.

link\movie 4.AVI

 

2. PDF files of publications:

Lux et al 2005.pdf: “Green fluorescent protein-tagged adeno-associated virus particles allow the study of cytosolic and nuclear trafficking”

Perabo and Endell et al 2006.pdf : “Combinatorial engineering of a gene therapy vector: directed evolution of adeno-associated virus”

 

3. PDF file of PhD thesis:

link\PhD thesis Lux.pdf

 

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