A biophysical assay for RNA based resistance

项目摘要 在抗击传染病方面，世界正迅速走向20世纪40年代前的局面。 疾病抗生素耐药性是全球范围内日益严重的问题，极大地阻碍了我们的 有能力平息全球流行病，如结核病和疟疾，以及简单的 肺部感染。该项目的意义重大，因为除非创新 开发了产生稳健和有效的新型抗生素的策略， 医疗保健费用将继续攀升，我们将完全失去打击 即使是最常见的感染。目前的抗生素治疗主要来自于 真菌和细菌产生的天然产物，能够抑制其他微生物的生长， 生物，通常通过抑制细胞壁的合成或维持，或通过抑制蛋白质 合成.自从1929年弗莱明首次分离出青霉素以来， 几代抗生素仍然非常类似于原始的天然产品，具有功能性 修改后的群体，以增加其在更广泛的病原体中的活性， 他们的副作用恶唑烷酮、糖肽类、β-内酰胺类和喹诺酮类显示出一些 未来的希望，但革兰氏阴性细菌感染仍然存在问题。 核酸是药物设计的有希望的途径，既作为治疗剂又作为靶点。这里 我们提出了一个创新的计划，用于识别一类新的配体， 特异于RNA元件，该RNA元件是抗生素抗性的重要因素， 几十种致病细菌菌株，我们提出了一种生物物理筛选试验， 用于鉴定这种配体。首先，如具体目标1中所述，我们将描述一个模型 已经商业合成的核酸结构域，其具有允许 结构和动力学性质的这种分子在本体溶液中。然后我们将合成 序列特异性RNA结合配体，并筛选这些靶向的缀合物文库， 序列特异性结合并抑制靶核酸（特异性目的2）。一 这种方法的成功应用将使我们能够沉默一类人的抵抗途径， 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.