Human Molecular Genetics Advance Access originally published online on April 27, 2005
Human Molecular Genetics 2005 14(12):1641-1650; doi:10.1093/hmg/ddi172
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In vitro and in vivo suppression of GJB2 expression by RNA interference
1Molecular Otolaryngology Research Laboratory, Department of Otolaryngology—Head and Neck Surgery, Interdepartmental Ph.D. Genetics Program, The University of Iowa, 200 Hawkins Drive—21151 PFP, Iowa City, IA 52242, USA and 2Department of Otolaryngology—Head and Neck Surgery, Okayama University Graduate School of Medicine, Okayama, Okayama 700-8558, Japan
* To whom correspondence should be addressed. Tel: +1 13193563612; Fax: +1 13193564108; Email: richard-smith{at}uiowa.edu
Received February 27, 2005; Revised April 12, 2005; Accepted April 22, 2005
Mutations in GJB2 (gap junction protein, beta-2) are the major cause of autosomal recessive non-syndromic hearing loss. A few allele variants of this gene also cause autosomal dominant non-syndromic hearing loss as a dominant-negative consequence of expression of the mutant protein. Allele-specific gene suppression by RNA interference (RNAi) is a potentially attractive strategy to prevent hearing loss caused by this mechanism. In this proof-of-principle study, we identified a potent GJB2-targeting short interfering RNA (siRNA) to post-transcriptionally silence the expression of the R75W allele variant of GJB2 in cultured mammalian cells. In a mouse model, this siRNA duplex selectively suppressed GJB2R75W expression by >70% of control levels, thereby preventing hearing loss. The level of endogenous murine Gjb2 expression was not affected. Our data show that RNAi can be used with specificity and efficiency in vivo to protect against hearing loss caused as a dominant-negative consequence of mutant gene expression.