Targeted Aptamer Probes for Nucleic Acid Detection

目前研究的最终目标是开发用于体内基因表达成像和体内靶向细胞杀伤或修复的探针。目前的提案描述了一种新的核酸探针设计，将为这些应用所需的新技术提供基础。这些探针也有望成为当前探针应用的重大改进，如分析食品中的细菌和其他污染，以及分析细胞和组织样本中的特定基因表达。我们称之为“靶向揭示适体探针”（TRAPS）。提出的TRAP设计的基本创新是包含一个衰减的适体，在与目标核酸杂交时显示。在目前的提案中，提出了以下具体目标：*产生一个基本的TRAP探针。确定当探针与互补RNA杂交时，TRAP探针仅与其配体结合。测试与探针结合的互补RNA和配体的数量之间的比例。检测TRAP与靶RNA相互作用的特异性。*生产捕获TRAP探针，可用于捕获分析并测试其有效性。建议的商业应用：最初，这些探针有望对目前用于检测DNA和RNA序列的核酸探针进行重大改进，例如分析细菌和其他食品污染以及分析细胞和组织样本中的特定基因表达。在进一步开发后，探针将用于基因表达的体内成像和靶向体内细胞，以便杀死或修复它们。

The eventual goal of the current research is to develop probes for in vivo imaging of gene expression and for in vivo targeting of cells for killing or remediation. The current proposal describes a novel nucleic acid probe design that will provide the basis of the new technology required for these applications. These probes are also expected to be a major improvement over current probes for applications such as the analysis of bacterial and other contamination of foods and in the analysis of specific gene expression in cell and tissue samples. We call these "Targeted Revealed Aptamer Probes" (TRAPS). The basic innovation in the proposed TRAP design is the inclusion of an attenuated aptamer that is revealed on hybridization with a target nucleic acid. In the current proposal, the following specific aims are proposed: * Produce a basic TRAP probe. Determine that the TRAP probe only binds its ligand when the probe is hybridized with complementary RNA. Test for proportionality between the amounts of complementary RNA and ligand binding to the probe. Test for specificity of the interaction between the TRAP and the target RNA. * Produce a capture TRAP probe that can be used in capture assays and test its efficacy. PROPOSED COMMERCIAL APPLICATIONS: Initially, these probes are expected to be a major improvement on the current nucleic acid probes used for detection of DNA and RNA sequences such as in the analysis of bacterial and other contamination of foods and in the analysis of specific gene expression in cell and tissue samples. After further development, the probes will be used for in vivo imaging of gene expression and for targeting cells in vivo so that they can be killed or remediated.