PNA HYBRIDIZATION OF FOLDED NUCLEIC ACID TARGETS

描述(改编自申请人的摘要)：本提案涉及 需要针对特定DNA和RNA序列的新试剂。是这样的 试剂在诊断和治疗方面都有潜在的用途 申请。建议中所描述的试剂是多肽核酸。 (PNA)低聚物，是DNA和RNA的合成类似物，其中 糖磷酸二酯主链被聚酰胺取代。PNA杂交到 具有高亲和力和高序列的互补单链DNA和RNA 选择性。PNA的大多数应用将需要与靶点杂交 序列是折叠结构的一部分，可能会施加显著的 通过严格阻止对目标站点的访问来阻止绑定。PNA可以 通过链入侵与双链DNA结合，但只有某些序列可以 有针对性的。人们对非双链DNA二级结构重要性的认识日益增强 结构和丰富多样的二级和三级结构 已经促使我们开始研究PNA靶向的能力 这些结构，无论是在简单模型系统中还是在 大的、折叠的RNA分子。我们将使用各种光谱， 量热法和凝胶电泳法研究热力学和凝胶电泳法 控制PNA与这些靶点杂交的动力学因素。校长 这项建议的目的是广泛地研究PNA杂交的范围 结构化目标，以更好地评估PNA在 诊断和治疗应用。以下具体目标将允许 我们要实现这一目标： *研究核酸与DNA和RNA发夹基序的结合。 *研究高嘧啶核糖核酸与单链和双链RNA的结合 序列 *评价PNA-DNA和PNA-RNA杂交动力学。 *RNA三级结构对PNA杂交的影响。

DESCRIPTION (adapted from applicant's abstract): This proposal addresses the need for new reagents targeted toward specific sequences of DNA and RNA. Such reagents would have potential use in both diagnostic and therapeutic applications. The reagents described (in the proposal) are peptide nucleic acid (PNA) oligomers, which are synthetic analogs of DNA and RNA in which the sugar-phosphodiester backbone is replaced by a polyamide. PNAs hybridize to complementary single-stranded DNA and RNA with high affinity and sequence selectivity. Most applications of PNA will require hybridization to a target sequence that is part of a folded structure that could impose significant barriers to binding by sterically blocking access to the target site. PNA can bind to duplex DNA by strand invasion but only certain sequences can be targeted. The growing awareness of the importance on nonduplex DNA secondary structures and the rich variety of secondary and tertiary structures exhibited by RNA have prompted us to initiate studies on the ability of PNA to target these structures, both in simple model systems and within the context of a large, folded RNA molecule. We will use a variety of spectroscopic, calorimetric and gel electrophoretic methods to study the thermodynamics and kinetic factors controlling PNA hybridization to these targets. The principal aim of this proposal is to broadly examine the scope of PNA hybridization to structured targets to better evaluate the potential utility of PNA in diagnostic and therapeutic applications. The following Specific Aims will allow us to achieve this goal: * Examine binding of PNA to DNA and RNA hairpin motifs. * Study binding of homopyrimidine PNA to single- and double-stranded RNA sequences * Evaluate PNA-DNA and PNA-RNA hybridization kinetics. * Effect of RNA tertiary structure on PNA hybridization.