Hi!
I thought it had always been accepted knowledge in the labs I've worked in that for a given promoter/protein pair, that IN GENERAL, you will get higher expression with transfection than with infection. This was in the context of lentiviral infection.
I'm helping a master's student write up her thesis, and now can't seem to find any sort of reference for this. Have I somehow misremembered or made this up, or do other people find this?
I understand of course that it will depend on the cell type and concentrations of the virus and the DNA etc etc. But leaving those aside - is there something to this? If so, is it also true for AAV or just lentivirus?
Thanks!
Jen
As far as I know, in my experience, virus infection yields much more efficient host gene transformation; therefore, potentially higher expression of manipulated genes. Viruses can enter/infect to lot more cells than transfecting plasmid DNA into the cells. Even though the transfection efficiency of plasmid DNA could be high, the probability of integration of that plasmid into the host cells' genome is still low. Although, with plasmid DNA one can get a good transient expression.
In my experiment with gene delivery (same plasmid DNA), lipofectamine 2000 and VSV-G pseudovirus were in use, we didn't get enough protein expression with lipofectamine 2000, the expression was so low that even with ECL the protein band was undetectable. This result has prompted us to another strategy, which was pseudovirus, the result was compared using flow cytometry, the later does yield a better result, which brought expression to about 10 times higher. But also, I wasn't able to quantify the lipofectamine nanoparticle, as same as pseudovirus.
Dear Jennifer Hazen
You have already mentioned the importance of cell type while describing your question and in my experience, this is the most important point to consider.
In my opinion selecting between transfection vs infection entirely depends on the target cell. The same also holds true between the choice of AAV vs lentiviral transduction.
For eg, cardiomyocytes are best transduced with AAVs (rather than plasmid transfection or lenti transduction) in our hands. For HEK cells all options work similarly for us based on our protocols.
So my answer here would be that the it all depends on the target cell that is being transfected/transduced and we cannot generalize which method is more efficient. We can however say which method is best suited for a specific cell type.
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