OLIGONUCLEOTIDE ANALOGS AS GENE REGULATION AGENTS

该计划项目的总体目标是开发耐核酸酶， 可用于控制基因的非离子寡核苷酸类似物 在mRNA或DNA水平以序列特异性方式表达。 修订后的申请包括以下项目：项目一， “甲基膦酸寡核苷的化学”，Paul S.米勒，P>I>; 项目二，“生理化学和分子细胞学”，保罗O。曹丕， 私家侦探，Stephen A. Lesko和简楼星博士，Co-P.I.;项目三， “作为抗病毒/抗癌剂的寡核苷酸类似物”，Laure博士 Aurelian，P.I.;和项目IV，“体外和体内肿瘤生长的调节 体内”，Esther Chang博士，P.I. 此外，还有两个核心组件， 科学核心，保罗·S。私家侦探米勒和行政核心， 博士保罗·O P. Ts'o，P.I. 在项目I中，新的化学方法包括 使用2 ′-O-甲基核糖核苷酸， 脱氧核糖核苷酸，手性（R和S）特异性Matagens的合成， 作为潜在前药的脱氧核糖核苷酸三酯的合成， 使用脱氧尿苷取代的低聚物，最后， 除了其它的与Matagen连接的新的烷基化剂之外， 用pesticen衍生的matagen的开发。 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-嘧啶三链体，以及 胞苷和各种类型的新型主链键。 调查 将包括光谱研究，NMR研究，保护分析， 量热法，但也将尝试与Matagen形成三链体 和活细胞中的目标DNA片段。 通过计算机 显微成像系统，单个RNA和DNA将可视化 用特定的探针定量。 在项目III中，Matagen和pavelien 衍生化的Matagen将用于研究速发型胶原的功能。 早期基因（IEmRNA 4.5编码IE68 K和IE 12 K）， 反式激活基因（IE110，IE 175），以及核糖核苷酸 还原酶基因（RR 1）。 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， Matagen和衍生的Matagen将用于抑制Ha-ras 癌基因表达（通过p21测量）与肿瘤相关性 在各种细胞和动物模型中的特性。

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.