DNA-based Arrays of Cross-reactive Molecular Sensors

描述（由申请人提供）：哺乳动物嗅觉系统由1000多个细胞上的1000个不同的受体组成，尽管单个受体可能只有适度的敏感性和选择性，但总的来说，它们能够通过平行的分子识别事件对痕量气味进行精细区分。相同受体的多个拷贝之间的相互作用和交叉反应受体的平行处理是当前感兴趣的领域。易受结合位点变化影响的人工传感器将允许研究平行识别作为分析物识别的策略。我们最近在三向和四向连接的固定框架内发现了具有疏水可变结构域的寡核苷酸，这使得我们能够构建此类研究所需的数千种不同的分子传感器。因此，将构建一组基于DNA的交叉反应疏水传感器来模拟嗅觉系统。在资助期结束时，一个典型的阵列将由一个到几百个选定的分子传感器组成，这些分子传感器被组织在1586孔板上，每个传感器有多个读数。该阵列将在收集的尿液样本上进行验证，用于代谢物筛选。疏水指纹将与疾病状态相关。该方法将首先用于库欣病、肾上腺腺瘤、肾上腺癌、多毛症和先天性肾上腺增生。在下一阶段，基于dna的低聚糖交叉反应传感器将被整合到阵列中，以扩大可在单步分析中分析的代谢物面板。

DESCRIPTION (provided by applicant): The mammalian olfactory system is composed of 1000 different receptors on over 1000 cells each, and although individual receptors may be only modestly sensitive and selective, in the aggregate they are capable of fine differentiation of trace odorants through parallel molecular recognition events The interplay between multiple copies of the same receptor and parallel processing of cross-reacting receptors is an area of current interest. Artificial sensors susceptible to facile variations of the binding site would permit investigation of parallel recognition as a strategy for analyte identification. Our recent discovery of oligonucleotides with a hydrophobic variable domain within the fixed frameworks of three- and four-way junctions allows construction of the several thousand distinct molecular sensors needed for such an investigation. Therefore, an array of cross-reacting hydrophobic sensors based on DNA will be constructed to model the olfactory system. The array will provide an instantaneous fingerprint of all molecules with hydrophobic domains present in a solution A typical array at the end of funding period will consist of one to several hundred selected molecular sensors organized in 1586-well plates with multiple readings per sensor. The arrays will be validated on urine specimens collected for metabolite screening. The hydrophobic fingerprints will be correlated with disease states. This approach will be initially demonstrated on Cushing's disease, adrenal adenoma, adrenal carcinoma, hirsutism and congenital adrenal hyperplasia. In the next phase, DNA-based cross-reactive sensors for oligosaccharides will be incorporated into arrays to expand the panel of metabolites that could be profiled in a single-step assay.