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Phoenix packaging cells and compatible plasmids


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#1 californiacricket

californiacricket

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Posted 23 October 2009 - 09:03 AM

I am looking to generate a siRNA retrovirus to transduce mouse cells with. I have cloned my shRNA into pLVTHM (second generation). The protocols all say that I need to co-transfect into 293T cells a gag-pol and env plasmid along with my siRNA containing plasmid (pLVTHM) that gets packaged. Can I use the Phoenix packaging cell line instead and just transfect the pLVTHM plasmid to get retrovirus? If not, why? I can't find any examples in the literature where people have used the pLVTHM plasmid with the Phoenix cell line. Also is there a good vector that I could clone my siRNA into and use the Phoenix cell line? Most of the vectors I have found are made to express a protein and don't have the shRNA promoters.




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