title: Efficient down-regulation of glia maturation factor expression in mouse brain and spinal cord.
abstract: 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.
authors.affiliation: Department of Neurology, The University of Iowa, 200 Hawkins Drive, Iowa City, IA 52242, USA.
authors.forename: Smita
authors.initials: S
authors.lastname: Zaheer
authors.forename: Yanghong
authors.initials: Y
authors.lastname: Wu
authors.forename: Xi
authors.initials: X
authors.lastname: Yang
authors.forename: Ramasamy
authors.initials: R
authors.lastname: Thangavel
authors.forename: Shailendra K
authors.initials: SK
authors.lastname: Sahu
authors.forename: Asgar
authors.initials: A
authors.lastname: Zaheer
mesh_terms: D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D015536:Down-Regulation; D004797:Enzyme-Linked Immunosorbent Assay; D020904:Glia Maturation Factor / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D034741:RNA, Small Interfering; D013116:Spinal Cord / Q000378:metabolism*
chemical_list: D020904:Glia Maturation Factor; D034741:RNA, Small Interfering