- 作者: Masashi Tanaka a,b, Harm-Jan Borgeld a,b, Jin Zhang a,b, Shin-ichi Muramatsu c, Jian-Sheng Gong a,b, Makoto Yoneda d, Wakako Maruyama a, Makoto Naoi b, Tohru Ibi f, Ko Sahashi f, Masayo Shamoto e, Noriyuki Fuku a,g, Miyuki Kurata a, Yoshiji Yamada a,b, Kumi Nishizawa a Yukihiro Akao a,b, Nobuko Ohishi b, Shigeaki Miyabayashi h, Hiraku Umemoto i, Tatsuo Muramatsu i, Koichi Furukawa j, Akihiko Kikuchi k, Imaharu Nakano c, Keiya Ozawa l, Kunio Yagi a,b
- 作者服務機構: a Department of Gene Therapy, Gifu International Institute of Biotechnology, b Institute of Applied Biochemistry, Mitake, c Department of Neurology, Jichi Medical School, Tochigi, d Second Department of internal Medicine, Fukui Medical University, Fukui, e Laboratory of Biochemistry and Metabolism, Departrnent of Basic Gerontology, National Institute for Longevity Sciences, Obu, f Neurologysection, Department of internal Medicine , Aichi Medical University, Aichi , g Japan Science and Technology Corporation, Tokyo, h Department of Pediatrics, Sendai National Hospital, Sendai, i Department of Applied Genetics and Physiology, Graduate School of Bioagricultural Sciences, Nagoya University, j Department of Biochemistry, Graduate school of Medical Sciences, Nagoya University, k Department of Medical Mycology, Graduate School of Medical Sciences, Nagoya University, Nagoya, l Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School, Tochigi, Japan
- 中文摘要: --
- 英文摘要: The restriction endonuclease Smal has been used for thediagnosis of neurogenic muscle weakness, ataxia andretinitis pigmentosa disease or Leigh's disease, causedby the Mt8993T→G mutation which results in aLeu156Arg replacement that blocks proton translocationactivity of subunit a of F F -ATPase. Our ultimate goal isto apply Smal to gene therapy for this disease, becausethe mutant mitochondrial DNA (mtDNA) coexists withthe wild-type mtDNA (heteroplasmy), and because onlythe mutant mtDNA, but not the wild-type mtDNA, isselectively restricted by the enzyme. For this purpose, wetransiently expressed the Smal gene fused to a mito-chondrial targeting sequence in cybrids carrying the mu-tant mtDNA. Here, we demonstrate that mitochondriatargeted by the Smal enzyme showed specific elimina-tion of the mutant mtDNA. This elimination was followedwith repopulation by the wild-type mtDNA, resulting inrestoration of both the normali ntracellular ATP level andnormal mitochondrial membranl potential. Further-more, in vivo electroporation of the plasmids expressingmitochondrion-targeted EcoRl induced a decrease in cy-tochrome c oxidase activity in hamster skeletal muscleswhile causing no degenerative changes in nuclei. Deliv-ery of restriction enzymes into mitochondria is a novelstrategy for gene therapy of a special form of mitochon-drial diseases.
- 中文關鍵字: --
- 英文關鍵字: Mitochondrial disease, Leigh's disease, Neurogenic muscle weakness, ataxia and retinitis pigmentosa, Gene therapy, Restriction endonuclease