title: Efficient Gene Suppression in Dorsal Root Ganglia and Spinal Cord Using Adeno-Associated Virus Vectors Encoding Short-Hairpin RNA.
abstract: RNA interference is a powerful tool used to induce loss-of-function phenotypes through post-transcriptional gene silencing. Small interfering RNA (siRNA) molecules have been used to target the central nervous system (CNS) and are expected to have clinical utility against refractory neurodegenerative diseases. However, siRNA is characterized by low transduction efficiency, insufficient inhibition of gene expression, and short duration of therapeutic effects, and is thus not ideal for treatment of neural tissues and diseases. To address these problems, viral delivery of short-hairpin RNA (shRNA) expression cassettes that support more efficient and long-lasting transduction into target tissues is expected to be a promising delivery tool. Various types of gene therapy vectors have been developed, such as adenovirus, adeno-associated virus (AAV), herpes simplex virus and lentivirus; however, AAV is particularly advantageous because of its relative lack of immunogenicity and lack of chromosomal integration. In human clinical trials, recombinant AAV vectors are relatively safe and well-tolerated. In particular, serotype 9 of AAV (AAV9) vectors show the highest tropism for neural tissue and can cross the blood-brain barrier, and we have shown that intrathecal delivery of AAV9 yields relatively high gene transduction into dorsal root ganglia or spinal cord. This chapter describes how to successfully use AAV vectors encoding shRNA in vivo, particularly for RNA interference in the central and peripheral nervous system.
authors.affiliation: Department of Orthopaedic and Spinal Surgery, Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan. enomoto.orth@tmd.ac.jp.|Hyperbaric Medical Center, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan. enomoto.orth@tmd.ac.jp.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Enomoto
authors.affiliation: Department of Orthopaedic and Spinal Surgery, Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan. hirai.orth@tmd.ac.jp.|School of Dentistry, Oral Biology, Oral Biology and Medicine, University of California Los Angeles, 10833 Le Conte Avenue, 63-078 CHS, Los Angeles, CA, 90095-1668, USA. hirai.orth@tmd.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Hirai
authors.affiliation: Department of Orthopaedic and Spinal Surgery, Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan. kaborth@tmd.ac.jp.
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Kaburagi
authors.affiliation: Department of Neurology and Neurological Science, Graduate School, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan. yokonuro@tmd.ac.jp.
authors.forename: Takanori
authors.initials: T
authors.lastname: Yokota
mesh_terms: D000818:Animals; D001522:Behavior, Animal; D001835:Body Weight / Q000235:genetics; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D005727:Ganglia, Spinal / Q000378:metabolism* / Q000821:virology; D055785:Gene Knockdown Techniques / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D051379:Mice; D008813:Mice, Inbred ICR; D035683:MicroRNAs / Q000235:genetics; D010641:Phenotype; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D013116:Spinal Cord / Q000378:metabolism* / Q000821:virology
chemical_list: D035683:MicroRNAs; D012333:RNA, Messenger; D034741:RNA, Small Interfering
keywords: Adeno-associated virus; Dorsal root ganglia; Intrathecal administration; RNA interference; Short-hairpin RNA; Spinal cord