Hey,
I heard several times in the lab that a plasmid performs better (higher transfection efficiency, more protein produced, longer present in cells, ...) when the plasmid is spliced.
(Meaning if your plasmid has the gene sequence with introns and exons, it would perform better compared to the plasmid with only the cDNA.)
The story goes that this improved performance is linked to the plasmid being processed by the splicing machinery.
Have you heard something similar? Do you know data supporting that?
My mean reason to be skeptic is that the plasmid becomes quite big due to all the introns. And usually you prefer having smaller plasmids to make transfection easier.
But it is true that it might "feel more natural" for a (eukaryotic) cell to have spliceable RNA...
Good question. Since splicing enhances translation (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC324426/), is it possible that you observe more transfected cells simply because you now also detect have less cells with low plasmid copy number?
Thank you Vesa! Very interesting paper.
Based on this paper you might almost wonder why we remove introns in our plasmids. Higher yield, correct localization of mRNA... all improved when you keep introns.
So I guess we use cDNA only out of convenience because less basepairs facilitate all our molecular protocols/techniques...
Whereas biology in the cell would rather speak in favor of using the whole gene...
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