Chemical Synthesis and Applications of Nucleic Acid Analogues

核酸类似物的化学合成及应用

• 批准号: RGPIN-2016-05126
• 负责人: Damha, Masad
• 金额: $ 6.63万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=708915
• 关键词: Chemical; Synthesis; Applications; Nucleic; Acid

Nucleic acids are the fundamental molecules of life, carrying the genetic blueprints for all living things on Earth. Refined through billions of years of evolution, they achieve an ideal balance between complexity and elegance, a perfect marriage of form and function. As such, synthetic DNA and RNA oligonucleotides and their analogues are incredibly useful tools for molecular biology, functional genomics, and biochemistry research. We will pursue three specific aims within this proposed NSERC Discovery Grant program. First, we will synthesize and characterize branched RNA analogues in order to study the structure and properties of the human lariat RNA debranching enzyme (hDbr1), the only enzyme known to hydrolyze 2',5'-phosphodiester bonds of lariat RNA introns; carry out structure-guided synthesis of non-hydrolyzable RNA branchpoint analogues through the introduction of sugar and base modifications to create conformations that mimic that of the lariat RNA branchpoint observed in nature; determine the structures of Dbr1inhibitor complexes, and test the toxicity of Dbr1 inhibitor compounds in neuronal cell lines. Second, we will design synthetically modified DNA and RNA structures that are able to "switch off" genes (e.g., disease-causing genes) by intercepting mRNA, by virtue of their expected increased binding affinity for RNA and stability in biological media. Third, we plan to study folded genomic DNA structures, i.e., the G-quadruplexes and i-motifs. While the former structures are already known to control gene expression, the biological role of i-motifs remains elusive. We propose to correlate, for the first time, telomerase activity with transient formation of G-quadruplexes and putative i-motifs within duplex DNA. The stabilization of i-motif structures at physiological conditions by our proposed approach will open new research directions on i-motifs and G-quadruplexes. Our proposed research program will capitalize on the expertise and research advances obtained during the previous granting period to further advance the state of the art and shed light on several clinically relevant questions in bioorganic chemistry. This proposed program minimizes risk through building on previous achievements, while still continuing our lab's theme of boldly pushing the leading edge of nucleic acids research. Our scientific approach combines solution and solid-phase synthetic methods, drug discovery, spectroscopic analysis, and molecular biology techniques. These studies expose my students to a wide range of topics in nucleic acid chemistry and biology research, and to strategies used for identifying candidate molecules and developing them into useful probes and/or therapeutic drugs.

核酸是生命的基本分子，携带着地球上所有生物的遗传蓝图。经过数十亿年的进化，它们在复杂性和优雅性之间达到了理想的平衡，是形式和功能的完美结合。因此，合成的DNA和RNA寡核苷酸及其类似物是分子生物学，功能基因组学和生物化学研究的非常有用的工具。 我们将在这个拟议的NSERC发现补助金计划中追求三个具体目标。首先，我们将合成和表征支链RNA类似物，以研究人IXRNA去分支酶（hDbr 1）的结构和性质，该酶是已知唯一水解IXRNA内含子的2 '，5'-磷酸二酯键的酶;进行结构导向的非可水解的RNA分支点类似物，通过引入糖和碱基修饰来产生模拟LIPO 2构象的构象在自然界中观察到的RNA分支点;确定Dbr 1抑制剂复合物的结构，并测试Dbr 1抑制剂化合物在神经元细胞系中的毒性。其次，我们将设计合成修饰的DNA和RNA结构，这些结构能够“关闭”基因（例如，致病基因），这是由于它们对RNA的预期增加的结合亲和力和在生物介质中的稳定性。第三，我们计划研究折叠的基因组DNA结构，即，G-四链体和i-基序。虽然已经知道前一种结构控制基因表达，但i基序的生物学作用仍然难以捉摸。我们建议关联，第一次，端粒酶活性与瞬时形成的G-四链体和假定的i-基序内双链体DNA。我们所提出的方法在生理条件下的i-模体结构的稳定性将开辟新的研究方向i-模体和G-四链体。 我们拟议的研究计划将利用在前一个资助期内获得的专业知识和研究进展，进一步推进最先进的技术水平，并阐明生物有机化学中的几个临床相关问题。该计划通过建立在以前的成就基础上，将风险降至最低，同时仍然继续我们实验室大胆推动核酸研究前沿的主题。 我们的科学方法结合了溶液和固相合成方法、药物发现、光谱分析和分子生物学技术。这些研究使我的学生接触到核酸化学和生物学研究的广泛主题，以及用于识别候选分子并将其开发为有用的探针和/或治疗药物的策略。