Journal
NEUROCHEMICAL RESEARCH
Volume 37, Issue 7, Pages 1578-1583Publisher
SPRINGER/PLENUM PUBLISHERS
DOI: 10.1007/s11064-012-0753-x
Keywords
Glia maturation factor (GMF); Short hairpin RNA (shRNA); Lentiviral vector; Gene silencing; Experimental autoimmune encephalomyelitis (EAE)
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Funding
- National Institute of Neurological Disorders and Stroke [NS073670, NS047145]
- VA Merit Review award
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Long-lasting siRNA-based down-regulation of gene of interest can be achieved by lentiviral-based expression vectors driving the production of short hairpin RNA (shRNA). We investigated an attractive therapeutic approach to target the expression of proinflammatory GMF by using lentiviral vector encoding GMF-specific shRNA to reduce GMF levels in the spinal cord and brain of mice. To determine the effect of GMF-shRNA on GMF protein levels, we performed quantitative ELISA analysis in brain and in thoracic, cervical and lumbar regions of spinal cord from mice followed by GMF-shRNA (G-shRNA) or control shRNA (C-shRNA) treatments. Our results show a marked reduction of GMF protein levels in brain and spinal cord of mice treated with GMF-shRNA compared to control shRNA treatment. Consistent with the GMF protein analysis, the immunohistochemical examination of the spinal cord sections of EAE mice treated with GMF-shRNA showed significantly reduced GMF-immunoreactivity. Thus, the down-regulation of GMF by GMF-shRNA was efficient and wide spread in CNS as evident by the significantly reduced levels of GMF protein in the brain and spinal cord of mice.
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