DEVELOPMENTS OF THE METHOD FOR THE CHEMICAL SYNTHESIS OF RNA CONTAINING MODIFIED BASED" -DIRECTED TOWARD FULL AUTOMATION OF RNA CONTAINING MODIFIED BASES BY USE OF NEW SYNTHETIC UNITS-

含有修饰碱基的 RNA 化学合成方法的发展”-通过使用新的合成单元实现含有修饰碱基的 RNA 的全自动合成-

基本信息

批准号：
07559005
负责人：
SEKINE Mitsuo
金额：
$ 9.79万
依托单位：
TOKYO INSTITUTE OF TECHNOLOGY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

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.

该项目的目的是开发新方法，以快速合成含有修饰碱基的RNA。我们首先成功地合成了U1 RNA的5'-末端寡聚核苷酸，该RNA在其5'-末端末端含有三甲基化帽的独特结构。 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'-将双磷酸化化学合成，用于在3'-末端位点的U1 RNA的5'-末端块的化学合成。2-氮杂氧化丙氨酸合成为核糖核苷模型，并将其与DNA寡素化合物合成。事实证明，事实证明，具有靶向互补的DNA链具有光链接能力。在这些研究中，我们显示了一种新的策略，即在不使用磷酸化的方法中使用新的冷凝试剂来使用磷酸化的碱性氧基核苷酸化学合成，而无需使用磷酸化的基础。在此DNA水平上获得了令人鼓舞的结果。在这些条件下，没有观察到基础座的侧反应。因此，我们现在将这种新策略扩展到化学合成改性RNA的化学策略，因为这种方法使我们能够合成含有2--寡脱氧核糖核苷酸的化学策略。叠氮化氧化丙氨酸和n-乙烯基氧基丁胺。