PEI transfection for lentivirus production

[Anthi Demetriadou]

Dear Julia,

in the Nat prot paper (Joung et al, 2017) it is recommended a low-cost protocol for lentivirus production using PEI (Box 5).

Specifically, it is suggested to use Polyethylenimine HCl MAX, Linear, Mw 40,000 (PEI Max; Polysciences, cat. no. 24765-1).

Do you know if I have to change something in the protocol if I use a different type of PEI (in the lab we have Polyethylenimine, Linear, MW 25000, Polysciences. cat. no. 23966)??

Thank you!

Anthi

[Mithun Das]

Hi Anthi,

Although you have asked Julia, I thought I should share my experience.

PEI 40,000 thousand is efficient than 25000, therefore, in a sense, you can increase the amount of PEI than what was used in Nature protocol, however, using a lot more PEI can be stressful to the cells. I guess you are going to quantify your lentivirus after production, therefore, you can try to use slightly modified method to get the best lentivirus titer.

Cheers

Mithun

[Anthi Demetriadou]

Thank you Mithum for sharing your knowledge!!

Anthi

[Julia Joung]

Hi all,

Mithun - thanks so much for sharing your expertise on using PEI with different MWs. I have never actually tested different versions of PEI, so would not have been able to advise Anthi.

Best,

Julia

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