Snap-To-It Probes

描述（由申请人提供）：微阵列提供了以高度复用格式描述遗传改变的潜力，但在目标捕获方面存在次优特异性。提高目标捕获特异性的阵列技术将通过允许制造具有优越性能特征的阵列而具有商业价值。我们建议开发一种新的Snap-To-It阵列技术，该技术由排列的PNA和DNA探针组成，这些探针受到分子内螯合物的构象约束。与靶标结合导致螯合物解离，探针与靶标核酸断裂，这是一个全有或全无的机制。与之前使用Snap-To-It探针进行的溶液阶段热力学研究一致，我们假设固定在阵列上的受限探针与无约束探针相比将表现出更好的目标特异性。所提出的螯合基序很容易引入到阵列使用现有的标准阵列合成方法，只有几个额外的工艺步骤。此外，螯合结合与沃森-克里克配对是正交的，因此不会干扰靶杂交。第一阶段演示了Snap-To-It探针的可行性，并为进入密集的第二阶段项目奠定了基础，该项目涉及开发和测试用于癌症分析的原型Snap-To-It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.