Team:Heidelberg/Project/Capsid Shuffling/Homology Based
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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. [https://2010.igem.org/Team:Heidelberg/Project/Mouse_Infection 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 (#43) 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. |
In addition, 12 viruses were packaged with a YFP construct and used to transduce primary hepatocytes, HeLa and Huh-7 cells. Flow cytometry measurements showed that some of those viruses (e.g. number 8) are even more efficient than many of the wildtype serotypes. However, the measurements will be repeated, since not enough time was allowed for the viruses to be produced and for the cells to express sufficient YFP for good measurements. | In addition, 12 viruses were packaged with a YFP construct and used to transduce primary hepatocytes, HeLa and Huh-7 cells. Flow cytometry measurements showed that some of those viruses (e.g. number 8) are even more efficient than many of the wildtype serotypes. However, the measurements will be repeated, since not enough time was allowed for the viruses to be produced and for the cells to express sufficient YFP for good measurements. | ||
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[[Image:facs_43_huh.gif|200px|clone number 43, Huh-7 cells]] | [[Image:facs_43_huh.gif|200px|clone number 43, Huh-7 cells]] | ||
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[[Image:hepg2_transyfp_2.gif|Clone 43 under microscope, HepG2 cells|300px]] | [[Image:hepg2_transyfp_2.gif|Clone 43 under microscope, HepG2 cells|300px]] | ||
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[[Image:huh7_transyfp1_2.gif|300px]] | [[Image:huh7_transyfp1_2.gif|300px]] | ||
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[[Image:Flowcyt.jpg|Flow cytometry result for the 12 clones as compared to the 12 wildtypes|300px]] | [[Image:Flowcyt.jpg|Flow cytometry result for the 12 clones as compared to the 12 wildtypes|300px]] | ||
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{{:Team:Heidelberg/Pagemiddle}} | {{:Team:Heidelberg/Pagemiddle}} | ||
[[Image:HomBasShuffling 934px.png|270px]] | [[Image:HomBasShuffling 934px.png|270px]] | ||
{{:Team:Heidelberg/Bottom}} | {{:Team:Heidelberg/Bottom}} |
Latest revision as of 03:59, 28 October 2010
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