TETHERED NUCLEOBASES FOR ENHANCED DNA RECOGNITION

用于增强 DNA 识别的束缚核碱基

• 批准号: 6460920
• 负责人: Steven C. Zimmerman
• 金额: $ 28.76万
• 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/6460920
• 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）本提案描述了一项重大的新 开发拴系DNA碱基的倡议。这些新基地由四个 标准的DNA碱基A、T、G和C，修饰后可容纳一个附件， 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。这些“超级基地” 应当能够以更高的亲和力与互补序列杂交 和选择性。它们还可以选择性地结合DNA碱基错配，提供 检测单核苷酸多态性（SNP）的机制。 每个新的DNA超级碱基的发展将遵循四个不同的步骤：（1） 合成.合成通常很短，从简单，可用的 材料.更复杂的基地被设计成后备基地， 如有必要，综合工作将需要更多的努力。(2)特征化。 除了建立分子结构外，重要的是检查 不希望的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它们的能力， 识别错配，我们将在有机溶剂中进行模型研究， 决定了氢键相互作用的质量。有利结合 是进一步研究和发展的基础。(4)承认 寡核苷酸超碱基将被掺入寡核苷酸中， 检测它们结合靶碱基的能力。二聚体超级碱基将是 合成，并将检查其识别错配的能力。 除了对这种承认的基本兴趣之外， 可以预见应用。其中包括使用超级基地， 治疗性寡核苷酸（例如，反义），SNP的检测，如 用于化学和分子生物学研究等的不可加工的双链体锚。