i have my gene of interest in this way(picture in attached files), Now I am confuse about whether i can use this whole thing to construct my one the 3 lentivirus packaging vector named pLenti vector,or I have to purify my fragment of interest out of this..kindly look at the attached files below,my gene of interest in interleukin 2 gene
Viral Production
The process of producing infectious transgenic lentivirus is outlined in the simple schematic to the right. 3-4 plasmids are transfected into A293T cells:
- one transfer plasmid
- one or two packaging plasmid(s)
- one envelope plasmid
After media change and a brief incubation period, supernatant containing the virus is removed and stored or centrifuged to concentrate virus. Crude or concentrated virus can then be used to transduce the cells of interest. For determination of viral titer and full details, see the (Link opens in a new window) protocol available at the (Link opens in a new window) Trono lab website.
For more detail, you can You can refer to this website.
https://www.addgene.org/guides/lentivirus/
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