The preparation and delivery of siRNA were performed as previously described with slight modification.32 (link) Briefly, Scramble RNA (Qiagen, 1022076) or myelin basic protein (MBP) siRNA (Qiagen, SI01302035) was diluted to 200 µM with RNase-free water, aliquoted, and stored at −80℃. Prior to administration, RNA/lipid complexes were prepared by mixing RNA (2 µg) with 8 µl i-Fect™ reagent (Neuromics). After incubating for 10 min at room temperature, 5 µL of the RNA/lipid complex was delivered into the lumbar region of the spinal cord by intrathecal injection. The injection was given daily for three consecutive days.
Protocol detail
Intrathecal Delivery of siRNA for Myelin Repair
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Corresponding Organization : The University of Texas Medical Branch at Galveston
Other organizations : Zhejiang Sci-Tech University
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Variable analysis
independent variables
- Scramble RNA (Qiagen, 1022076)
- Myelin basic protein (MBP) siRNA (Qiagen, SI01302035)
- Outcome of the treatment (not explicitly mentioned)
- RNase-free water
- I-Fect™ reagent (Neuromics)
- Injection volume (5 µL)
- Injection location (lumbar region of the spinal cord)
- Injection frequency (daily for three consecutive days)
- Scramble RNA (Qiagen, 1022076)
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