Over these past several years, I have communicated our progress of working with customers on in vivo and in vitro transfection in neurons and neuronal cell lines using ouri-FectTMsiRNA Transfection Kit.
First presented at SFN in 2004, I as pleased to highlight a recent publication in Molecular Pain demonstrating the inhibition of a GPCR expression in vivo in spinal cord and DRG via intrathecal delivery of siRNA. I would also like to thank Dr. Luo for her excellent technical work in developing the detailed Protocols Summaries used to deliver the siRNA with i-Fect.
An Efficient Intrathecal Delivery of Small Interfering RNA to the Spinal Cord and Peripheral Neurons.
Luo MC, Zhang DQ, Ma SW, Huang YY, Shuster SJ, Porreca F, Lai J.
We have developed a highly effective method for in vivo gene silencing in the spinal cord and dorsal root ganglia (DRG) by a cationic lipid facilitated delivery of synthetic, small interfering RNA (siRNA). A siRNA to the delta opioid receptor (DOR), or a mismatch RNA, was mixed with the transfection reagent, i-FectTM (vehicle), and delivered as repeated daily bolus doses (0.5 microgram to 4 micrograms) via implanted intrathecal catheter to the lumbar spinal cord of rats. Twenty-four hours after the last injection, rats were tested for antinociception by the DOR selective agonist, [D-Ala2,Glu4]deltorphin II (DELT), or the mu opioid receptor (MOR) selective agonist, [D-Ala2, N-Me-Phe4, Gly-ol5]enkephalin (DAMGO). Pretreatment with the siRNA, but not the mismatch RNA or vehicle alone, blocked DELT antinociception dose-dependently. The latter was concomitant with a reduction in the spinal immunoreactivity and receptor density of DOR, and in DOR transcripts in the lumbar DRG and spinal dorsal horn. Neither siRNA nor mismatch RNA pretreatment altered spinal immunoreactivity of MOR or antinociception by spinal DAMGO, and had no effect on the baseline thermal nociceptive threshold. The inhibition of function and expression of DOR by siRNA was reversed by 72 hr after the last RNA injection. The uptake of fluorescence-tagged siRNA was detected in both DRG and spinal cord. The low effective dose of siRNA/i-FectTM complex reflects an efficient delivery of the siRNA to peripheral and spinal neurons, produces no behavioral signs of toxicity, and this delivery method may be optimized for other gene targets.
Here's a complete listing of our Transfection Reagents-Kits Publications.
Also check out our transfection methods and data blog: http://sirnatransfection.blogspot.com/