Intrathecal Delivery of siRNA
Another publication referencing successful delivery of siRNA using i-Fect
Suneeta Tumati, Tally Largent Milnes, Henry I. Yamamura, Todd W. Vanderah, William R. Roeske and Eva V. Varga. Intrathecal Raf-1-selective siRNA attenuates sustained morphine-mediated thermal hyperalgesia. doi:10.1016/j.ejphar.2008.10.033
...siRNAs stock solutions (100 ?M) were prepared in double distilled RNAse free water and stored in aliquots at ?80 °C. For intrathecal treatment, aliquots of the stock solution (2 ?g of the appropriate siRNA) were mixed (1:5 v/v)with i-Fect transfection reagent (Neuromics, Edina, MN). After recovery from the surgery (5-7 days), the animals received intrathecal injections (2 ug! siRNA/1 0 ul/rat) of either a lipid encapsulated Raf-1-selective siRNA mixture (Smart pool siRNA, Dharmacon Inc; Chicago, IL, Cat # L-087699-00) (Raf-1 siRNA groups) or i-Fect encapsulated non-targeting dsRNA (Dharmacon, #D-001810-01-20) (control mismatch siRNA groups) or the transfection lipid alone, once daily, for 3 days, as described earlier (Gardell et al., 2002). Intrathecal injections of the siRNAs or the transfection agent alone did not cause any sign of behavioral toxicity. Western blots, using a Raf-1-selective antibody, indicated that intrathecal treatment with the Raf-1-selective siRNA mixture for 3 days significantly reduced Raf-1 protein levels in the dorsal root ganglion and in the dorsal horn of the spinal cord..
Lasani S. Wijetunge, Sally M. Till, Thomas H. Gillingwater, Cali A. Ingham, and Peter C. Kind. mGluR5 Regulates Glutamate-Dependent Development of the Mouse Somatosensory Cortex. The Journal of Neuroscience, December 3, 2008, 28(49):13028-13037; doi:10.1523/JNEUROSCI.2600-08.2008.
...Western blotting was performed as mentioned above and membranes were probed with antibodies against mGluR5 (1:4000, Neuromics)...
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