Is polyA signal necessary for gene expression in lentiviral vectors?

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Kristoffer Riecken

Polyadenylation is done by the vector backbone, the polyA signal is located within the 3'-LTR. Don't clone an additional polyA signal into the vector, this would severely reduce the titer! http://tronolab.epfl.ch/lentivectors

James Sturges

Hello, I have a question. Could a lentiviral vector escape detection in a vaccine, such as a flu shot? If so, under what circumstances? The reason I am asking is for a work of fiction, so speculation is quite welcome. Thank you! ------------Regards, James

Suparerk Borwornpinyo

I want to add some information on the polyA signal. It becomes necessary to put the polyA signal following your gene of interest. Particularly we need a very strong one for sufficient termination, thus preventing the read-through into cellular genes. It is an optimal design for safety gene transfer using integrating vectors.

Hello. Could someone please tell me if my question is too ill-informed to be answered ... or what? A little feedback from you specialists would be much appreciated. Is this topic taboo? Thanks. ------------Regards, James

Govinda Sharma

What about if I wanted to express two separate transgenes under the control of two separate promoters? Something like 5'LTR-prom1-CDS1-prom2-CDS2-3'LTR. Would I need to include a polyA signal after CDS1?

Also, is this the best orientation to express the two coding regions? I've been told by others that transcribing one of the cds from the other strand would be a good way to prevent crosstalk, but if I'm relying on the 3' LTR to provide polyA signal, then the cds that's transcribed in the opposite direction wouldn't be going through the 3' LTR.

Josephine F Trott

James Sturges, you would do best to ask a vaccine manufacturer what quality control tests are routinely done on vaccines. If your'e talking about sabotage and a work of fiction, then of course anything is possible. But it's very easy to detect minute traces of any DNA sequence (which lentiviruses contain) using PCR.