I'm setting up an assay to weed out compounds that inhibit transfection efficiency? Are there any known small molecule compounds that affect transfection efficiency that I can use as a control rather than just a mock transfection control?
Hi Elizatbeth,
I can only offer you my best guess.
Considering that transfection is based on liposomes composed of cationic phospholipids, any surfactant will disturb/disrupt the liposomes even at very low concentration. More over a non anionic (Triton X100) or anionic surfactant (SDS) will also interfere with the charges of the liposomes.
You could also use anionic phospholipids which will decrease the binding of the liposomes with the negatively charged genetic material (that you want to transfect).
More generally, anything lipophylic will interfere with liposome formation; anything that mask/neutralize lipid charges will interfere with DNA-liposomes association and thereby transfection.
Well...those are wild assumptions.
Hi,
On cell culture level we have seen with peptide-based transfection systems that scavenger receptor inhibitors/blockers work by almost completely blocking the transfection.
Have a look: (1) Ezzat, K., Helmfors, H., Tudoran, O., Juks, C., Lindberg, S., Padari, K., El-Andaloussi, S., Pooga, M., and Langel, U. (2012) Scavenger receptor-mediated uptake of cell-penetrating peptide nanocomplexes with oligonucleotides. FASEB J. 26, 1172–80.
You can find some autophagy related small molecules here:
Article Role of autophagy in cell-penetrating peptide transfection model
It should be noted that there are reports about scavenger receptors being recruited to the cell surface from cytoplasm upon transfection. Therefore they might not be your small molecule target.
Cheers
Hi, as pointed out by Tonis after Cyrille's answer, there is not only lipid-based reagents... I would suggest using endocytosis inhibitors/blockers, it works for all chemical transfectants. Best, Sandra
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