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Nucleic Acids Research Advance Access published online on June 25, 2008
Nucleic Acids Research, doi:10.1093/nar/gkn371
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© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

Efficient delivery of RNA Interference to peripheral neurons in vivo using herpes simplex virus

Anna-Maria Anesti1,*, Pieter J. Peeters2, Ines Royaux2 and Robert S. Coffin1

1BioVex Inc., 34 Commerce Way, Woburn, MA 01801, USA and 2Johnson and Johnson Pharmaceutical Research and Development, Turnhoutseweg 30, B-2340 Beerse, Belgium

*To whom correspondence should be addressed. Tel: +1 781 376 4900; Fax: +1 781 933 6025; Email: manesti{at}biovex.com

Received February 18, 2008. Revised May 14, 2008. Accepted May 28, 2008.

Considerable interest has been focused on inducing RNA interference (RNAi) in neurons to study gene function and identify new targets for disease intervention. Although small interfering RNAs (siRNAs) have been used to silence genes in neurons, in vivo delivery of RNAi remains a major challenge limiting its applications. We have developed a highly efficient method for in vivo gene silencing in dorsal root ganglia (DRG) using replication-defective herpes simplex viral (HSV-1) vectors. HSV-mediated delivery of short-hairpin RNA (shRNA) targeting reporter genes resulted in highly effective and specific silencing in neuronal and non-neuronal cells in culture and in the DRG of mice in vivo including in a transgenic mouse model. We further establish proof of concept by demonstrating in vivo silencing of the endogenous trpv1 gene. These data are the first to show silencing in DRG neurons in vivo by vector-mediated delivery of shRNA. Our results support the utility of HSV vectors for gene silencing in peripheral neurons and the potential application of this technology to the study of nociceptive processes and in pain gene target validation studies.


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