| Project/Area Number |
07559005
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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 |
広領域
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| Research Institution | TOKYO INSTITUTE OF TECHNOLOGY |
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Principal Investigator |
SEKINE Mitsuo TOKYO INSTITUTE OF TECHNOLOGY,FACULTY OF BIOSCIENCE AND BIOTECHNOLOGY,ASSOCIATE PROFESSOR, 生命理工学部, 助教授 (40111679)
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| Co-Investigator(Kenkyū-buntansha) |
KIDA Masahide WAKO PURE CHEMICAL LTD., CENTRAL INSTITUTE OF WAKO PURE CHEMICAL,CHIEF RESEARCHE, 大阪中央研究所, 首席研究員
ISHIKAWA Masahide TOKYO INSTITUTE OF TECHNOLOGY,GRADUATE SCHOOL,DEPARTMENT OF INTELLIGENCE SCIENCE, 総合理工学研究科, 助手 (50212858)
WADA Takeshi TOKYO INSTITUTE OF TECHNOLOGY,FACULTY OF BIOSCIENCE AND BIOTECHNOLOGY,DEPARTMENT, 生命理工学部, 助手 (90240548)
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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 |
¥15,300,000 (Direct Cost: ¥15,300,000)
Fiscal Year 1996: ¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1995: ¥9,300,000 (Direct Cost: ¥9,300,000)
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| Keywords | ANTISENCE NUCLEIC ACID RNA / MODIFIED RNA / SYNTHETIC / TRANSFER RNA SYNTHESIS / PHOTOCROSSLINKING / SOLID PHASE / 2-AZIDODEOXYADENOSINE / U1 RNA / RNA / 4-チオウリジン / 6-チオイノシン / 6-チオグアノシン / アミダイト法 / ホスホネート法 / 縮合反応 |
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| Research Abstract |
THE PURPOSE OF THIS PROJECT IS IS TO DEVELOPE NEW METHODS FOR THE RAPID SYNTHESIS OF RNA CONTAINING MODIFIED BASES.WE FIRST SUCCEEDED IN SYNTHESIZING THE 5'-TERMINAL OLIGORIBONUCLEOTIDES OF U1 RNA CONTAINING A UNIQUE STRUCTURE OF TRIMETHYLATED CAP AT ITS 5'-TERMINAL END.WE HAVE ALSO ESTABLISHED PHOSPHORAMIDITE BUILDING BLOCKS OF 6-THIOGUANOSINE,6-THIOINOSINE,AND 4-THIOURIDINE AS NEW PHOTOCROSSLINKING SITES.THESE MODIFIED RIBONUCLEOSIDES COULD BE INCORPORATED INTO RNA BY USING THESE SYNTHETIC UNITS.PARTICULARLY,4-THIOURIDINE 3', 5'-BISPHOSPHATE WAS CHEMICALY SYNTHESIZED AND USED FOR THE CHEMICAL SYNTHESIS OF THE 5'-TERMINAL BLOCK OF U1 RNA CONTAINING 4-THIOURIDINE AT THE 3'-TERMINAL SITE.2-AZIDODEOXYADENOSINE WAS SYNTHESIZED AS AN RIBONUCLEOSIDE MODEL AND INCORPORTED INTO DNA OLIGOMERS.THESE MODIFIED OLIGOMERS.HAVE PROVED TO HAVE A PHOTOCROSSLINKING ABILITY WITH TARGETED COMPLEMENTARY DNA STRANDS.IN ADDITION TO THESE STUDIES,WE HAVE SHOWED A NEW STRATEGY OF THE CHEMICAL SYNTHESIS OF OLIGODEOXYRIBONUCLEOTIDES WITHOUT USING BASE PROTECTING GROUPS BY THE PHOSPHONATE APPROACH USING A NEW CONDENSING REAGENT,BOMP.VERY PROMISING RESULTS WERE OBTAINED AT THIS DNA LEVEL.NO SIDE REACTIONS ON THE BASE MOITIES WERE OBSERVED UNDER THESE CONDITIONS.THEREFORE,WE ARE NOW EXTENDING THIS NEW STRATEGY TO THE CHEMICAL SYNTHESIS OF MODIFIED RNA,SINCE THIS APPROACH ENABLED US TO SYNTHESIZE OLIGODEOXYRIBONUCLEOTIDES CONTAINING 2-AZIDODEOXYADENOSINE AND N-ACETYLDEOXYCYTIDINE.
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