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Transient and permanent gene transfer into the brain of the teleost fish medaka (Oryzias latipes) using human adenovirus and the Cre-loxP system
Authors:Yuji Suehiro  Teruhiro Okuyama  Kiyoshi Naruse  Takeo Kubo  Hideaki Takeuchi
Institution:a Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan
b Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
c Laboratory of BioResource, National Institute for Basic Biology, Myodaiji, Okazaki, Aichi 444-8585, Japan
d Hashimoto Research Unit, Brain Science Institute, RIKEN, Wako, Saitama 351-0198, Japan
Abstract:In this study, we demonstrated that human type-5 adenovirus infected the brain of the teleost fish, medaka (Oryzias latipes), in vivo. Injection of adenoviral vector into the mesencephalic ventricle of medaka larvae induced the expression of reporter genes in some parts of the telencephalon, the periventricular area of the mesencephalon and diencephalon, and the cerebellum. Additionally, the Cre-loxP system works in medaka brains using transgenic medaka carrying a vector containing DsRed2, flanked by loxP sites under control of the β-actin promoter and downstream promoterless enhanced green fluorescent protein (EGFP). We demonstrated that the presence of green fluorescence depended on injection of adenoviral vector expressing the Cre gene and confirmed that EGFP mRNA was transcribed in the virus-injected larvae.
Keywords:Medaka  Conditional gene manipulation  Brain
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