DEALING WITH ANTIBIOTIC RESISTANCE--ANTISENSE TECHNOLOGY

描述(摘自申请者摘要)：抗药性是一个主要原因 这是克服细菌感染的障碍。这项提案针对的是 细菌感染性疾病治疗中的关键问题：需要 制定旨在保持现有的有效性的战略 氨基糖苷类抗生素。调查人员关注的是对 氨基糖苷6‘-N-乙酰基转移酶介导的氨基糖苷阿米卡星 Aac(6‘)-Ib，它还可以修饰其他几种氨基糖苷类抗生素。这个 这项研究项目的长期目标是开发反义的使用 寡核苷酸选择性抑制AAC(6‘)-Ib的表达 和其他相关基因。这项赠款提案的具体目标是：1) 促进切割的人工合成寡核苷酸及其类似物的鉴定 核糖核酸酶H或核糖核酸酶P.1b对AAc(6‘)-Ib mRNA的影响 合成寡核苷酸和寡核苷酸类似物。RNase H或RNase P 可以通过降解细胞中的mRNA来介导基因表达的抑制 存在合适的反义分子。这方面的总体战略是 具体目标将包括使用两个概念上不同的 确定信使核糖核酸中可与之相互作用区域的方法 寡核苷酸。有了这些信息，寡脱氧核苷酸及其类似物 将通过诱导核糖核酸酶H抑制AAC(6‘)-Ib的表达 信使核糖核酸降解。这些信息还将用于 抑制血管内皮细胞生长因子表达的反义寡核苷酸的研究进展 AAc(6‘)-Ib通过诱导核糖核酸酶P降解mRNA。接下来， 研究人员将启动对细胞摄取寡核苷酸和 类似物。2a)抑制的肽核酸(PNA)分子的发展 AAC(6‘)-Ib的表达。2)PNA分子的生物利用度研究。一本小说 使用反义技术抑制基因表达的策略现已可用 与合成多肽核酸(PNA)有关。PNA可以开发为 原核系统中的抗菌剂。调查人员将测试内部 几种PNA分子的体内外干扰活性 AAC(6‘)-Ib的表达。然后，研究人员将研究生物利用度。 裸露的和脂质体包裹的PNA分子。

DESCRIPTION (Adapted from Applicant's Abstract): Drug resistance is a major obstacle in the conquest of bacterial infections. This proposal targets a critical issue in the treatment of bacterial infectious diseases: the need to develop strategies aimed at preserving the effectiveness of currently available aminoglycoside antibiotics. The investigators focus on resistance to the aminoglycoside amikacin mediated by the aminoglycoside 6'-N-acetyltransferase AAC(6')-Ib, which can also modify several other aminoglycoside antibiotics. The long term goal of this research project is to develop the use of antisense oligonucleotides as tools to selectively inhibit the expression of aac(6')-Ib and other related genes. The specific aims for this grant proposal are: 1a) Identification of synthetic oligonucleotides and analogs that promote cleavage of aac(6')-Ib mRNA by RNase H or RNase P. 1b) Bioavailability studies of synthetic oligonucleotides and oligonucleotide analogues. RNase H or RNase P can mediate inhibition of gene expression by degradation of mRNA in the presence of appropriate antisense molecules. The general strategy of this specific aim will consist of the utilization of two conceptually different approaches to identify regions in the mRNA available for interaction with oligonucleotides. With this information, oligodeoxynucleotides and analogues will be designed to inhibit the expression of aac(6')-Ib by inducing RNase H degradation of the mRNA. The information will also be utilized for the development of antisense oligoribonucleotides that inhibit the expression of aac(6')-Ib by inducing RNase P degradation of the mRNA. Following, the investigators will initiate studies on the cell uptake of oligonucleotides and analogues. 2a) Development of peptide nucleic acid (PNA) molecules that inhibit expression of AAC(6')-Ib. 2b) Bioavailability studies of PNA molecules. A novel strategy to inhibit gene expression using antisense technology is now available with the synthesis of peptide nucleic acids (PNAs). PNAs may be developed as antimicrobial agents in prokaryotic systems. The investigators will test the in vitro and in vivo activity of several PNA molecules to interfere with the expression of aac(6')-Ib. The investigators will then study the bioavailability of naked and liposome encapsulated PNA molecules.