Can I make lentivirus with pLXSN retrovirus plasmid?

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vmurthy
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Can I make lentivirus with pLXSN retrovirus plasmid?

Post by vmurthy » Thu Sep 07, 2017 6:14 pm

I am trying to make lentivirus with retroviral pLXSN (I read that pLXSN can be used for both lenti and retrovirus).

I would prefer to use the 2nd generation lentiviral packaging system with pMD2.G and psPAX2 in 293T cells. Is this possible? I have looked for papers that use this system/ use LXSN for lentivirus production, but haven't been able to find any. If anyone has come across any papers with this, please let me know.

Plasmid maps:
pLXSN: https://www.addgene.org/vector-database/3492/
pMD2.G: https://www.addgene.org/12259/
psPAX2: https://www.addgene.org/12260/

Thanks!

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Vmedvil
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Post by Vmedvil » Wed Sep 27, 2017 5:24 am

No, you cannot make a functioning Retroviral/Lentiviral Vector(Which are actually the same Retroviruses are a class of Lentiviruses). You have problems kinda the pLXSN Plasmid is missing the Pol gene only having Gag, Both Pol and Gag are required for a functioning Vector, but it could be used as a Lentiviral Vector if you find one with Pol and VSV-G in it. The psPAX2 Plasmid has both HIV-1 Pol and Gag in it, so it can construct the full DNA machinery of the Vector. pMD2.G has VSV-G which is the Envelope of the Vector in it, you would need to put both pMD2.G and psPAX2 plasmids into 293T cells to produce a Fully functioning Lentiviral Vector, but the pLXSN Plasmid is missing the Pol gene of the Virus. Secondly, you are not currently transferring any DNA via the Vector without DNA which has LTR3' and LTR5' Primers around it. These Vectors if you make them Pol-Gag-VSV-G will not able to make more of themselves after infection without the Primers around the Pol-Gag-VSV-G Genes, I would suggest not including them around them, much less dangerous, but you should include them around the genes you are sending into the target cells max length 10,000 base pairs.

Genes needed for Lentiviral Vector
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