Design of Hammerhead Ribozymes that Cleave Murine Sry mRNA in vitro and in vivo
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概要
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As the first step in investigating the possiblity of applying ribozyme technology to artificial control of the sex ratios at birth in farm animals, where the demand for females exceeds that for males, we designed a hammerhead ribozyme (HHRz) and 2 tRNAval-hammerhead ribozyme complexes (tRNARz3 and tRNARz4), and examined their effects upon murine Sry mRNA in vitro and in cells. We demonstrated that HHRz and tRNARz3 could effectively cleave the target Sry mRNA in vitro. For the purpose of experiments in vivo, HHRz was cloned into the highly efficient pUC-CAGGS mammalian expression vector (pCAG/HHRz), and the tRNA ribozyme complexes were cloned into the pol III promoter-driven pPUR-KE vector (pPUR/tRNARz3 and pPUR/tRNARz4); the ribozyme vectors were co-transfected with the target vector (pCAG/Sry). A suppressive action (up to approx. 60%) was confirmed for pCAG/HHRz and pPUR/tRNARz3 upon the transiently expressed exogenously introduced Sry in M15 cultured cells.
著者
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NISHINO Koichiro
Laboratory of Applied Genetics, Graduate School of Agricultural and Life Sciences, University of Tok
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Tachi Chikashi
Laboratory of Applied Genetics, Department of Animal Resource Science, School of Agriculture and Lif
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SONOKI Shigenori
Laboratory of Environmental Chemistry, Faculty of Environment and Health, Azabu University
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ITO Masanori
Gene Function Research Center, National Institute of Advanced Industrial Science and Technology (AIS
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Hisamatsu Shin
Laboratory Of Environmental Chemistry College Of Environmental Health Azabu University
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Taira Kazunari
Gene Discovery Research Center National Institute Of Advanced Industrial Sciences And Technology (ai
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NISHINO Koichiro
Laboratory of Cellular Biochemistry, Department of Animal Resource Sciences, School of Agriculture and Life Sciences, The University of Tokyo
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TACHI Chikashi
Laboratory of Develomental and Reproductive Biotechnology, Department of Animal Resource Sciences, School of Veterinary Medicine and Life Sciences, Azabu University
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KAWASAKI Hiroaki
Gene Function Research Center, National Institute of Advanced Industrial Science and Technology
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IWAMORI Tokuko
Laboratory of Develomental and Reproductive Biotechnology, Department of Animal Resource Sciences, School of Veterinary Medicine and Life Sciences, Azabu University
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IWAMORI Tokuko
Laboratory of Applied Genetics, Graduate School of Agricultural and Life Sciences, The University of Tokyo
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HISAMATSU Shin
Laboratory of Environmental Chemistry, School of Environmental Health, Azabu University
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NAKAMURA Kaori
Laboratory of Develomental and Reproductive Biotechnology, Department of Animal Resource Sciences, School of Veterinary Medicine and Life Sciences, Azabu University
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MURATA Chisato
Laboratory of Develomental and Reproductive Biotechnology, Department of Animal Resource Sciences, School of Veterinary Medicine and Life Sciences, Azabu University
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NISHIMURA Seiichiro
Laboratory of Develomental and Reproductive Biotechnology, Department of Animal Resource Sciences, School of Veterinary Medicine and Life Sciences, Azabu University
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NAKAYAMA Aya
Gene Function Research Center, National Institute of Advanced Industrial Science and Technology
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SUYAMA Eigo
Gene Function Research Center, National Institute of Advanced Industrial Science and Technology
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