TETHERED NUCLEOBASES FOR ENHANCED DNA RECOGNITION

用于增强 DNA 识别的束缚核碱基

• 批准号: 6623068
• 负责人: Steven C. Zimmerman
• 金额: $ 29.8万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6623068
• 关键词: X ray crystallography; adenine; antisense nucleic acid; complementary DNA; cytosine; guanosine; hydrogen bond; nuclear magnetic resonance spectroscopy; nucleic acid chemical synthesis; nucleic acid purification; nucleic acid structure; nucleobase; nucleobase analog; oligonucleotides; thymine

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.

描述：（由申请人提供）该提案描述了一项重大新内容 主动开发束缚DNA碱基。这些新基地包括四个 标准 DNA 碱基 A、T、G 和 C，经过修改以容纳能够 额外的氢键（超出沃森克里克）和碱基堆积 与互补 DNA 链的相互作用。因此，这些“超级基地” 应该能够以更高的亲和力杂交互补序列 和选择性。它们还可以选择性地结合 DNA 碱基错配，从而提供 检测单核苷酸多态性（SNP）的机制。 每个新 DNA 超级碱基的开发将遵循四个不同的步骤：(1) 合成。合成通常很短并且从简单、可用的开始 材料。更复杂的基地被设计作为备份和他们的 如果需要的话，合成将需要更多的努力。 (2)表征。 除了确定分子结构外，检查分子结构也很重要 不良互变异构平衡的可能性。 (3)模型研究。之前 将超级碱基插入寡核苷酸或测试其能力 认识到不匹配，我们将在有机溶剂中进行模型研究 确定氢键相互作用的质量。良好的结合力 为进一步学习和发展奠定了基础。 (4) 认可 寡核苷酸。超级碱基将被整合到寡核苷酸中并 研究了它们结合目标碱基的能力。二聚体超级碱基将是 合成并检查它们识别不匹配的能力。 除了这种类型的认可的根本利益之外，各种 可以设想应用。其中包括使用超级基地 治疗性寡核苷酸（例如反义）、SNP 检测，如 用于化学和分子生物学研究等的不可加工的双链锚。