TETHERED NUCLEOBASES FOR ENHANCED DNA RECOGNITION

描述：(由申请人提供)本提案描述了一项重大的新 发展系留DNA碱基的倡议。这些新基地由四个 标准DNA碱基A、T、G和C，经修饰后可容纳能够 额外的氢键(超越沃森-克里克)和碱基堆积 与互补DNA链的相互作用。因此，这些“超级基地” 应该能够与具有更高亲和力的互补序列杂交 和选择性。它们还可能选择性地结合DNA碱基错配，从而提供 单核苷酸多态(SNPs)检测机制。 每个新的DNA超级碱基的开发将遵循四个不同的步骤：(1) 综合。这些合成一般都很短，而且从简单的、可用的开始 材料。更复杂的基地被设计为后备基地，他们的 如果需要，合成将需要更多的努力。(2)人物塑造。 除了建立分子结构，重要的是要检查 不理想的互变异构体平衡的可能性。(3)模型研究。在.之前 在寡核苷酸中插入超碱基或测试它们的能力 识别错配，我们将在有机溶剂中进行模型研究，以 确定氢键相互作用的质量。有利的捆绑 为进一步研究和开发奠定了基础。(4)承认 寡核苷酸。超级碱基将被纳入寡核苷酸和 他们结合目标碱基的能力被检查。二聚超碱基将是 并检查它们识别错配的能力。 除了对这种类型的认识的根本利益之外，还有各种 应用是可以预见的。其中包括使用超级碱基在 治疗性寡核苷酸(例如反义)、SNPs的检测、AS 不可加工的双链锚定，用于化学和分子生物学研究等。

DESCRIPTION: (provided by applicant) This proposal describes a major new initiative to develop tethered DNA bases. These new bases consist of the four standard DNA bases A, T, G, and C, modified to hold an appendage capable of additional hydrogen bonding (beyond Watson-Crick) and base-stacking interactions with the complementary DNA strand. As such, these "super bases" should be capable of hybridizing complementary sequences with higher affinity and selectivity. They may also selectively bind DNA base mismatches providing a mechanism for detecting single nucleotide polymorphisms (SNPs). The development of each new DNA super-base will follow four distinct steps: (1) Synthesis. The syntheses are generally short and start with simple, available materials. More sophisticated bases have been designed as back-ups and their synthesis, if needed, will require somewhat more effort. (2) Characterization. Beyond establishing the molecular structure, it is important to examine the possibility of undesirable tautomeric equilibria. (3) Model Studies. Prior to inserting super-bases into oligonucleotides or testing their ability to recognize mismatches, we will perform model studies in an organic solvent to determine the quality of the hydrogen bonding interactions. Favorable binding qualifies a base for further study and development. (4) Recognition of oligonucleotides. Super-bases will be incorporated into oligonucleotides and their ability to bind target bases examined. Dimeric super-bases will be synthesized and their ability to recognize mismatches will be examined. Beyond the fundamental interest in this type of recognition various applications can be envisioned. These include the use of super-bases in therapeutic oligonucleotides (e.g., antisense), detection of SNPs, as nonprocessable duplex anchors for chemical and molecular biology studies, etc.