siRNA transfection of primary neurons

DanSolo via methods%40net.bio.net (by daniel.otoole At ucd.ie)
Fri Nov 24 06:13:22 EST 2006


O'Hare wrote:
> Has anybody managed to successfully transfect primary cultures of
> cerebellar granule neurons with short double stranded siRNA?  If so,
> what transfection reagent did you use and how much RNA and transfection
> reagent needs to be combined to achieve optimal transfection?  Any
> additional details that could be provided would also be greatly
> appreciated.  Thank you.
>
We tried this with primary rat neurons/astrocytes/co-cultures, and
nothing would get any nucleotide in. Lipofectin, Lipofectamine 2000,
Genjuice etc. We ended up electroporating, though 50%+ of the cells
die. I'm afraid I can't tell you how this will work for siRNA because
we were using luciferase reporter constructs.
Have you though of making a lentiviral vector for it? Once it's made
you can pretty much bet on it working on every cell type, and in vivo.



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