Sequence-Specific Incorporation of Functionalized Bases into RNA

这一关于“序列特异性掺入”研究的建议 将碱基功能化为RNA”。瑞士和 Steven A.本纳 位于瑞士苏黎世的Hochschule由NSF赞助。 核糖核苷酸参与细胞分裂，遗传调节， 病毒生长和肿瘤转化。 最近发现的 催化RNA及其作为RNA聚合酶的能力， 恢复了早期的想法，即第一个生物催化剂是 RNA分子。 关于这个问题的信息不多- 这些分子在溶液中的运动。 RNA的研究 RNA等活性中三级和四级结构 剪接，tRNA成熟和翻译受到限制， 现有RNA合成技术缺乏灵活性， nologies。 含有结构探针的非天然碱基（照片- 亲和标记，自旋标记），如果在特定位点掺入， 将促进这些领域的进步。 一种核苷酸碱基对 与A/T（U）和G/C不同， 通过聚合酶可以允许这样的序列特异性引入 结构探针 本工作的目标是合成异胞苷碱基 在5-位携带官能团，并将它们 在DNA模板的指导下酶促转化为RNA 含有d-iso-G。 iso-C/iso-G碱基对具有Watson-Crick 与A/T（U）和G/C碱基不同的氢键模式 对. 由于这种不同的氢键模式， 聚合酶应该忠实地掺入异C衍生物 只与d-iso-G相反。 这些功能化的碱将有助于 用于探测RNA的三级和四级结构。

This proposal for research on "Sequence Specific Incorporation of Functionalized Bases into RNA" between Dr. Christopher Y. Switzer and Professor Steven A. Benner of the Eidgenoessische Technische Hochschule, Zurich, Switzerland, is sponsored by NSF. Ribonucleotides are involved in cell division, genetic regulation, viral growth, and tumor transformation. The recent discovery of catalytic RNA and its ability to serve as an RNA polymerase has resurrected early ideas that the first biological catalysts were RNA molecules. There is little information regarding the confor- mation of these molecules in solution. The investigation of RNA tertiary and quaternary structure in such activities as RNA splicing, tRNA maturation, and translation has been limited by the lack of flexibility available with existing RNA synthesis tech- nologies. Unnatural bases containing structural probes (photo- affinity labels, spin-labels), if incorporated at specific sites, would facilitate advances in these areas. A nucleotide base pair distinct from those of A/T(U) and G/C that is incorporated into RNA by a polymerase could allow such sequence-specific introduction of structural probes. The goal of the proposed work is to synthesize the iso-cytidine bases carrying functional groups at the 5-position, and to incorporate them enzymatically into RNA under the direction of a DNA template containing d-iso-G. The iso-C/iso-G base pair has a Watson-Crick hydrogen bonding pattern different from the A/T(U) and G/C base pairs. Because of this different hydrogen bonding pattern, a RNA polymerase should faithfully incorporate the iso-C derivatives opposite only to d-iso-G. These functionalized bases will assist in experiments to probe RNA tertiary and quaternary structure.