DNA-based Arrays of Cross-reactive Molecular Sensors

描述（申请人提供）：哺乳动物的嗅觉系统由1000个不同的受体组成，每个受体上有1000多个细胞，尽管单个受体可能只有适度的敏感性和选择性，在聚合物中，它们能够通过平行的分子识别事件精细区分痕量气味剂。反应受体是当前感兴趣的领域。 对结合位点的容易变化敏感的人工传感器将允许调查平行识别作为分析物识别的策略。我们最近发现的寡核苷酸与疏水可变结构域内的固定框架的三路和四路路口允许建设的几千个不同的分子传感器需要这样的调查。因此，基于DNA的交叉反应疏水传感器阵列将被构建来模拟嗅觉系统。该阵列将提供溶液中存在的具有疏水结构域的所有分子的瞬时指纹图谱。在资助期结束时，典型的阵列将由组织在1586孔板中的一到几百个选定的分子传感器组成，每个传感器具有多个读数。 将在采集用于代谢物筛查的尿液样本上验证阵列。 疏水指纹将与疾病状态相关。 这种方法将首先证明库欣病，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的寡糖交叉反应传感器将被整合到阵列中，以扩大可以在单步测定中进行分析的代谢物组。

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.