Does anyone have experience transfecting or electroporating linear dsDNA into HEK cells? If yes what is the size limit, what size have you tried and worked.
In principle, I want to do HDR, and was thinking of delivering my replacement gene as a dsDNA rather than in a plasmid.
thank you
Dharanija
I have done plasmid transfection in HEK293T cells with polyethylenimine (PEI), a stable cationic polymer and transfection efficiency is very good . Did not done with dsDNA.
This is certainly possible in terms of delivering the DNA to the cells both electroporation and lipofectamine will work. The questions however why are you trying to deliver the DNA fragment into your cell. DNA by itself definitely not going to incorporate itself into the cell genome or express any protein as it requires more complex machinery such as promoters that are not present on your strand, what you will most probably get is a virus-like infection response from the cells as the reaction for the intracellular presence of the foreign DNA.
I've had success using the PiggyBac transposon system & electroporation with multiple mammalian cell lines, including HEK cells.
- Badges
- Science method