| Project/Area Number |
07557078
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Hematology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
HATTORI Toshio Kyoto University Ins.Vira Res Associate Professor, ウイルス研究所, 助教授 (30172935)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMAYAMA Takashi Hitachi Chemical Co.Ltd.Ass.Resarcher, 筑波開発研究所, 研究員補
YAMADA Ryo Hitachi Chemical Co.Ltd.Chief Resarcher, 筑波開発研究所, 主任研究員
UCHIYAMA Takashi Kyoto Univer.Jng.Virus Pea.Professor, ウイルス研究所, 教授 (80151900)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 1996: ¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1995: ¥8,000,000 (Direct Cost: ¥8,000,000)
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| Keywords | HIV / Ribozyme / RNA |
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| Research Abstract |
The ribozyme (Rz) was syntheisizes which recognizes relatively conserved sequences in neutralizing epitope (V3 loop) of envelope protein of HIV-1. Wild tye Rz and phosphorothioate DNA-RNA chimeric ribozyme (thio-Rz) were synthesized. Rz was labeled with fluorescence isithiocyanate and was introduced to cells with transfectam. After delivery into the cells, both wild type and thio-Rz were able to be readily detected. The positive signals faded within 12 hr in the former case, while they lasted 24 hr in the latter case. The thio-Rz as well as wild type ribozyme cleaved RNA transcripts (414 base) of V3 region, and produced the expected products (236 base and 178 base). We have developed a new assay system for envelope medaited fusion assay in which DiI labeled SupT1 cells were co-cultured with gp120 expressing cells (CHO-WT). In this assay, the fusion was associated with a dye transfer to CHO cells. The inhibitory effect by thio-Rz on cell fusion between envelope expressing CHO cells and CD4^+ cells appeared to be time-dependent, and about 70% of inhibition was seen at 72 hr. The mechanisms for inhibition of fusion ishould be clarified.
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