OLIGONUCLEOTIDE ANALOGS AS GENE REGULATION AGENTS

该计划项目的总体目标是开发抗核酸酶， 可用于基因调控的非离子寡核苷酸类似物 以序列特异性的方式在mRNA或DNA水平上表达。 本修订申请由以下项目组成：项目一、 《低聚核苷甲基膦酸盐的化学》，保罗·S·米勒； 项目II，“生理化学和分子细胞学”，Paul O.P.To‘o博士， P.I.，Stephen A.Lesko博士和Lou-sing Kan博士，共同P.I.；项目III， “作为抗病毒/抗癌药物的寡核苷酸类似物”，Laure博士 和项目IV，“肿瘤在体外和体内生长的调节” 此外，还有两个核心组件， 科学核心、私人助理保罗·S·米勒博士和行政核心， Paul O.P.To博士，P.I.在项目I中，新的化学方法包括 2‘-O-甲基核糖核苷酸的使用 脱氧核糖核苷酸，手性(R和S)特异性基质的合成， 潜在前体药物脱氧核糖核酸三酯的合成 脱氧尿苷取代低聚物的应用及发展 与Matgen连接的新型烷基化试剂 补骨脂素衍生化成熟剂的研究进展。在项目II中，研究 三链体的形成将包括嘧啶-嘧啶-嘌呤 三链体和嘌呤-嘌呤-吡啶三链体，以及类似物 胞苷和各种类型的新型主链连接。这项调查 将包括光谱研究、核磁共振研究、保护分析和 量热法，但也将尝试与Matgen形成三联体 以及活细胞中DNA的目标片段。通过计算机 显微成像系统，单个RNA和DNA将被可视化 用特定的探针进行定量。在项目III中，马特根和补骨脂素 衍生的基质将被用来研究即刻的功能 早期基因(编码IE68K和IE12K的IEmRNA4.5)，调节 反式激活基因(IE110、IE175)以及核糖核苷酸 还原酶基因(RR1)。这些研究对于理解 神经细胞中的病毒反式激活和病毒生长。在项目IV中， 基质和衍生基质将被用来抑制Ha-ras 癌基因表达(用p21衡量)与肿瘤的相关性 在各种细胞和动物模型中的性质。

The overall goal of this Program Project is to develop nuclease resistant, nonionic oligonucleotide analogs which can be used to control gene expression in a sequence specific manner at either the mRNA or DNA level. This revised application consists of the following projects: Project I, "Chemistry of Oligonucleoside Methylphosphonates", Paul S. Miller, P>I>; Project II, "Physiochemistry and Molecular Cytology", Dr. Paul O. P. Ts'o, P.I., Dr. Stephen A. Lesko and Dr. Lou-sing Kan, Co-P.I.; Project III, "Oligonucleotide Analogs as Antiviral/Anticancer Agents", Dr. Laure Aurelian, P.I.; and Project IV, "Modulation of Tumor Growth In Vitro and In Vivo", Dr. Esther Chang, P.I. In addition there are two core components, the Scientific Core, Dr. Paul S. Miller, P.I. and the Administrative Core, Dr. Paul O. P. Ts'o, P.I. In Project I, new chemical approaches include the use of 2'-O-methylribose nucleotides in addition to deoxyribonucleotides, the synthesis of chiral (R and S) specific Matagens, the synthesis of deoxyribonucleotide triesters as potential pro-drugs, the use of deoxyuridine substituted oligomers and finally, the development of novel alkylating agents attached to Matagen in addition to the further development of matagen derivatized with psoralen. In Project II, the study of triplex formation will include both pyrimidine-pyrimidine-purine triplexes and purine-purine-pyrimideine triplexes, as well as analogs of cytidine and various types of novel backbone linkages. The investigation will include spectroscopic studies, NMR studies, protection assay, and calorimetry, but also attempt will be made to form triplex with the Matagen and the target segment of DNA in living cells. Through the computer microscopic imaging system, individual RNA and DNA will be visualized quantitatively with specific probes. In Project III, Matagen and psoralen derivatized Matagen will be used to investigate the function of immediate early genes (IEmRNA 4.5 encoding IE68K and IE12K), the regulatory transactivating genes (IE110, IE175), as well as the ribonucleotides reductase gene (RR1). These studies are important to the understanding of viral transactivation and virus growth in neural cells. In Project IV, Matagen and derivatized Matagen will be used to suppress the Ha-ras oncogene expression, (as measured by the p21) in correlation to neoplastic properties in various cellular and animal models.