A biophysical assay for RNA based resistance

项目总结 在抗击传染性疾病方面，世界正迅速走向1940年前的S情景 疾病。抗菌素耐药性是全球范围内一个日益严重的问题，极大地阻碍了我们的 平息结核病和疟疾等全球流行病的能力，以及简单的 葡萄球菌感染。拟议的项目意义重大，因为除非具有创新性 制定了生产强健有效的新型抗生素的策略， 医疗费用将继续攀升，我们将完全失去抗击能力 即使是最常见的感染。目前的抗生素治疗主要源于 由真菌和细菌产生的能够抑制其他细菌生长的天然产物 生物体，通常通过抑制细胞壁的合成或维持，或通过抑制蛋白质 综合。自从1929年弗莱明首次分离出青霉素以来，随后的大多数 几代抗生素仍然与原始的天然产品非常相似，具有功能性 修改后的组在更广泛的病原体范围内提高其活性，并降低 他们的副作用档案。恶唑烷酮、糖肽、β-内酰胺类和喹诺酮类药物显示出一些 前景看好，但革兰氏阴性细菌感染仍然存在问题。 核酸是药物设计的很有前途的途径，既是治疗药物，也是靶点。这里 我们提出了一种创新的计划来鉴定一类新的配体 对一种RNA元件具有特异性，该元件是引起抗生素耐药性的重要因素 几十个致病菌菌株，我们提出了一种生物物理筛选方法 用于识别这样的配体。首先，正如具体目标1中所概述的，我们将描述一个模型 已经商业合成的核酸结构域，其修饰允许 该分子在本体溶液中的结构和动力学性质。然后我们将合成 序列特异的RNA结合配体，并筛选这些靶向的连接物文库 序列特异性地结合和抑制目标核酸(特定目的2)。一个 这种方法的成功应用将允许我们让一类人的抗性途径静默 被广泛使用的抗生素--氨基糖苷类。

PROJECT SUMMARY The world is rapidly heading towards a pre-1940's scenario when it comes to fighting infectious disease. Antimicrobial resistance is a growing problem on a global scale, greatly hampering our abilities to quell worldwide epidemics such as tuberculosis and malaria, as well as the simple staphylococcus infection. The proposed project is significant because unless innovative strategies are developed to produce robust and effective new classes of antibiotics, health care costs will continue to climb and we will completely lose our ability to combat even the most common infection. Current antibiotic treatments originated predominantly from natural products produced by fungi and bacteria that were able to inhibit the growth of other organisms, usually by inhibiting cell wall synthesis or maintenance or by inhibiting protein synthesis. Since penicillin was first isolated by Fleming in 1929, most of the subsequent generations of antibiotics remain very similar to the original natural products, with functional groups modified to increase their activity across a broader range of pathogens and decrease their side effect profiles. Oxazolidones, glycopeptides, b-lactams, and quinolones show some promise for the future, but gram-negative bacterial infections still remain problematic. Nucleic acids are promising avenues for drug design, both as therapeutics and as targets. Here we propose an innovative plan for identification of a novel class of ligands that are specific for an RNA element that is an important factor in the antibiotic resistance in dozens of pathogenic bacterial strains, and we propose a biophysical screening assay for identifying such ligands. First, as outlined in Specific Aim 1, we will characterize a model nucleic acid domain that has been synthesized commercially with modifications allowing structural and dynamic properties of this molecule in bulk solution. We will then synthesize sequence-specific RNA binding ligands and screen these targeted library of conjugates for sequence-specifically binding and inhibiting the target nucleic acid to (Specific Aim 2). A successful application of the approach will allow us to silence the resistance pathway for a class of widely used antibiotics-the aminoglycosides.