| Project/Area Number |
06454252
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
内科学一般
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| Research Institution | St.Marianna University School of Medicine |
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Principal Investigator |
SHIMADA Jingoro St.Marianna University School of Medicine Professor, 医学部, 教授 (50056701)
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| Co-Investigator(Kenkyū-buntansha) |
SHOJI Yoko St.Marianna University School of Medicine Assistant Professor, 医学部, 助手 (00235714)
MIZUSHIMA Yutaka Institute of Medical science, St.Marianna University School of Medicine Professo, 難病治療研究センサ, 教授 (40010409)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 1996: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1995: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1994: ¥2,900,000 (Direct Cost: ¥2,900,000)
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| Keywords | antisense DNA / phosphorothioate / cationic liposomes / cellular distribution / geraniol / ゲラニオール / カチオン性リポソーム / リポフェクチン / DOTAP / transfectam / VEGF / HUVEC / S-オリゴ / FITC標識S-オリゴ / 拡散チャンバー / 単純ヘルペスウイルスI型 / S-oligo / 共焦点レーザー蛍光顕微鏡システム |
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| Research Abstract |
Summary
We synthesized antisense DNA toward to immediate early pre-mRNA 4/5 (IE pre-mRNA4/5) of herpes simplex virus type1 (HSV-1) and evaluated anti-herpetic activity both in vitro and in vivo system. We clarified sequence specificity of antisense DNA,membrane delivery effects, and enhancement of biological activity by using drug delivery system (DDS). Since sequence non-specific activity of antisense DNA could not be neglected, anti-HIV activity of antisense DNA containing consecutive G sequence were evaluated.
We observed from this study following points ; 1) Phosphorothioate oligonucleotide possessed most potent anti-herpetic activity, which was paralleled with intercellular amount. 2) Cellular distribution of antisense DNA in infected cells were obviously different from that in non-infected cells. From this results, it was suggested that virus infection may enhance the delivery efficiency of antisense DNA.3) We found that antisense DNA targeted limited splicing site revealed potent anti-herpetic activity. 4) Sequence specific activity of antisense DNA can be delineated from calculation based on the thermodynamics and kinetics of base-pairing. 5) 5'end modification with geraniol enhanced anti-herpetic activity of antisense DNA in a sequence specific manncr. Homogeneous cellular distribution of geranil modified antisense DNA may reflect on anti-herpetic activity. 6) Cationic liposomes altered subcellular distribution of antisense DNA.7) Sequence non-specific biological activity was recognized. Sequence containing consecutive G sequence showed potent anti-HIV activity. 8) Inhibition of virus replication by antisense DNA was recognized on rabbit herpes cornea keratitis.
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