SYNTHESIS AND USE OF PHOSPHORUS MODIFIED OLIGONUCLEOTIDE

磷修饰寡核苷酸的合成及用途

• 批准号: 3497718
• 负责人: RICHARD H TULLIS
• 金额: $ 5万
• 依托单位: IMMUNE COMPLEX CORPORATION
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-06-15 至 1986-12-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3497718
• 关键词: DNA; complementary DNA; gel electrophoresis; genetic manipulation; genetic translation; messenger RNA; nucleic acid chemical synthesis; nucleic acid inhibitor; nucleic acid sequence; oligonucleotides; radiopharmacology; synthetic nucleic acid; tissue /cell culture

DNA hybridization reactions, discovered and explored in the early 1960s and 70's, exhibit nearly absolute specificity. Researchers recognized that this specificity might make it possible to develop DNA based diagnostic and therapeutic agents. In 1977 Paterson and coworkers discovered hybrid-arrest of translation. Anti-sense oligonucleotides, RNAs and cDNAs have now been shown to hybridize to and inhibit the translation of specific mRNAs both in vitro and in vivo. Such specific translation inhibiting compounds are useful in studying the function of particular mRNAs in the living cell and could potentially be used to selectively block the growth of viruses and bacteria. The major difficulty is that they are large polyanions which are poorly taken up by cells and are subject to rapid degradation by nucleases. Synthetic oligonucleotide phosphotriesters (PTEs) and methylphosphonates are non-ionic, lipophilic molecules which are stable to nucleases and well taken up by living cells. DNA phosphotriesters hybridize to both RNA and DNA in vitro and can selectively inhibit in vitro translation. Thus defined sequence PTE's would be useful in the study of mRNA function and processing in vivo. In addition, these agents could, if proven safe and effective, be used as specific and designable antiviral and antibiotic agents.

DNA杂交反应，在20世纪60年代初被发现和探索， 70年代，表现出几乎绝对的特异性。 研究人员认识到， 这种特异性可能使开发基于DNA的诊断和 治疗剂。 1977年帕特森和同事发现 翻译的混合逮捕。 反义寡核苷酸、RNA和cDNA 已经显示出与特异性DNA杂交并抑制其翻译， 在体外和体内的mRNA。 这种特异性翻译抑制 化合物可用于研究特定mRNA在细胞中的功能， 活细胞，并有可能用于选择性地阻止生长 病毒和细菌。 最大的困难是， 聚阴离子很少被细胞吸收， 通过核酸酶降解。 合成寡核苷酸磷酸三酯（PTE）和甲基膦酸酯 是非离子亲脂分子，对核酸酶稳定， 被活细胞吸收 DNA磷酸三酯与RNA杂交， 并能选择性地抑制体外翻译。 因此 确定的序列PTE在mRNA功能的研究中是有用的， 体内加工 此外，如果这些药物被证明安全， 有效，用作特异性和可设计的抗病毒和抗生素 剂.