I want to silence a gene in primary cells using siRNA approach. I have already seen effects of miRNA mimics in similar cell line but on different gene by adding them directly into media (7 days culture). I am not sure if the approach will work with siRNA. I am interested in suggestions if someone has done this without any vehicle (transfection reagent).
Naked short double stranded RNAs do not do great without a delivery vehicle mainly due to stability and poor uptake. Having said that, if the siRNA is chemically modified then it may work (which may explain what you got with the mimic). If the 5' sense strand is tagged with short peptide or cholesterol then the uptake would be higher.
Thanks Hasan,
I am also thinking little bit same. Now, can you suggest any company making peptide tagged LNA based siRNA.
Thanks again
There are two methods: reverse and direct. Try using Lipofectamine along with reverse transfection
I would also suggest tio use lipofection of short RNAs to treansfect these cells at collagen coated inserts.
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