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lentiviral systems for overexpression: clone in full cDNA or ORF? - (Aug/20/2008 )

I use lentiviral packaging system from Open Biosystems and am about to make some new constructs for gene overexpression. While ordering cDNA clones from Origene, a tech support specialist told me that if I am going to use lentiviral systems I am better of cloning in ORFs rather than the full lengths cDNAs with 5' and 3' regions because polyA tail on the clone may inhibit lentiviral packaging. I have never come across this bit of information and can not find any references on-line to support or disprove it. I am just a numbnut lentiviral system user and know very little about their biology. Does anyone have any input on this topic?
Thank you very much,
Christine

-Lapa-

Never heard of that neither, but still maybe possible.

I would go with the ORF, unless you have evidence the UTR regions could play a role in the protein's expression in some way (mRNA stability, traductability, etc)

-Madrius-

QUOTE (Madrius @ Aug 21 2008, 10:54 AM)
Never heard of that neither, but still maybe possible.

I would go with the ORF, unless you have evidence the UTR regions could play a role in the protein's expression in some way (mRNA stability, traductability, etc)


Thank you Madrius!
I am putting in ORFs and will try one construct with and without UTRs just in case.
christine.

-Lapa-

Some proteins are expressed better with the UTR's. That's protein dependent. Also poly A is not used in lentiviral vectors that we have or our collaborators.

-scolix-