lentiviral system: cloning strategy - (Oct/19/2009 )

Hi everyone,
I’m currently thinking in some strategies to clone a cDNA in a lentiviral vector system. Usually when I did this in other common expression mammalian systems (pCDNA 3, for example) I added the Kozak sequence just prior my insert observing the correct frame. However, a friend of mine that uses the lentiviral system said to me this is unnecessary and that he usually amplifies a small part of the 5’UTR (around 50 bp) together with his insert. According to him this is sufficient to get a good expression of the transgene. What do I do? I’m afraid get bad results in gene expression by using my friend’s strategy…
are there someone expert on this matter??

Depending on which lenti vector you're going to use, some lentiviral vectors give very good expression and you might say "no Kozak" is still working. However Kozak sequence definitely helps your CDS expression.

Thanks a lot for your words!!

Perhaps it is better to add this sequence instead assume the risk of getting bad expression levels with my vector, which is a non commercial one (obtained from one of our collaborators).

Functional Screens on Oct 20 2009, 04:06 PM said: