Team:Heidelberg/Project/Capsid Shuffling/Homology Based
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'''Selection in Huh-7 cells:''' | '''Selection in Huh-7 cells:''' | ||
- | Three selection rounds were carried out on Huh-7 cells, the viral DNA was isolated for each selection round and cloned into a Wilson AAV8 wild-type vector. 12 clones from each selection round were picked, and viruses that contain a YFP construct were produced and used to infect 96-well plates of mouse primary hepatocytes, HeLa and Huh-7 cells. Flow cytometry measurements | + | Three selection rounds were carried out on Huh-7 cells, the viral DNA was isolated for each selection round and cloned into a Wilson AAV8 wild-type vector. 12 clones from each selection round were picked, and viruses that contain a YFP construct were produced and used to infect 96-well plates of mouse primary hepatocytes, HeLa and Huh-7 cells. Flow cytometry measurements are to follow! |
'''Selection in Mouse Primary Hepatocytes:''' | '''Selection in Mouse Primary Hepatocytes:''' | ||
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'''Randomly picked clones''' | '''Randomly picked clones''' | ||
- | In parallel, we picked 48 randomly selected clones from the AAV library, packaged them with a YFP construct and tested them using flow cytometry on HepG2, Huh-7 and mouse primary hepatocytes. One clone of those was able to infect both HepG2 and Huh-7 cells, and was thus packaged with a luciferase construct and injected into mice. Mice experiment showed a specific targeting into the liver. | + | In parallel, we picked 48 randomly selected clones from the AAV library, packaged them with a YFP construct and tested them using flow cytometry on HepG2, Huh-7 and mouse primary hepatocytes. One clone of those was able to infect both HepG2 and Huh-7 cells, and was thus packaged with a luciferase construct and injected into mice. [https://2010.igem.org/Team:Heidelberg/Project/Mouse_Infection Mice experiment] showed a specific targeting into the liver. |
[[Image:hepg2_transyfp_1.tif|350px]] | [[Image:hepg2_transyfp_1.tif|350px]] |
Revision as of 02:47, 28 October 2010
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