Snap-To-It Probes

描述(由申请人提供)：微阵列提供了以高度多元化的形式描述基因改变的潜力，但受到靶标捕获的次优特异性的影响。通过制造具有优异性能特征的阵列，提高目标捕获的特异性的阵列技术将具有商业价值。我们建议开发一种新的Snap-to-It阵列技术，它由排列的PNA和DNA探针组成，这些探针受到分子内螯合物的构象限制。与靶的结合会导致螯合解离，探针以一种要么全有要么全不的机制与靶核酸结合。与以前使用Snap-to-It探针进行的溶液相热力学研究一致，我们假设固定在阵列上的限制性探针将比非限制性探针表现出更好的靶向性。使用现有的标准阵列合成方法，只需几个额外的工艺步骤，就可以很容易地将所提出的螯合基序引入阵列中。此外，螯合结合与Watson-Crick配对是正交的，因此不会干扰靶标杂交。第一阶段证明了Snap-to-It探针的可行性，并为进入密集的第二阶段计划奠定了基础，该计划涉及用于癌症分析的Snap-to-It原型阵列的开发和测试。 溶液和高通量微阵列中的DNA探针是用于从临床诊断到基础研究的无价工具。我们提出的Snap-to-It探针技术将产生具有卓越性能特征的微阵列。

DESCRIPTION (provided by applicant): Microarrays offer the potential to profile genetic alterations in a highly multiplexed format, but have suffered from sub-optimal specificity of target capture. Array technologies which improve the specificity of target capture will be of commercial value by allowing the manufacture of arrays with superior performance characteristics. We propose to develop a novel Snap-To-It array technology that consists of arrayed PNA and DNA probes that are conformationally constrained by an intra-molecular chelate. Binding to target results in the chelate dissociation and the probe snapping to the target nucleic acid in what is an all-or-none mechanism. Consistent with previous, solution phase, thermodynamic studies using Snap-To-It probes, we hypothesize that constrained probes immobilized on an array will exhibit superior target specificity compared to unconstrained probes. The proposed chelate motifs are easily introduced into arrays using existing standard array synthesis methods with only a few additional process steps. Further, chelate binding is orthogonal to Watson-Crick pairing, and thus will not interfere with target hybridization. Phase I demonstrated feasibility of Snap-To-It probes and set the stage to move into an intensive Phase II program involving the development and testing of prototype Snap-To-It arrays for cancer analysis. DNA probes in solution and in high-throughput microarrays are invaluable tools used in areas from clinical diagnostics to fundamental research. The Snap-To-It probe technology we propose will produce microarrays with superior performance characteristics.