RNA Aptamers Recognizing RNA Structural Elements

9319139施密特最近的研究表明，核酸分子可以折叠成各种形状，能够与小分子或蛋白质配体发生特定的相互作用。这项申请建议从包含大约10个(Ex13)随机序列的DNA模板库中合成复杂的RNA物种种群。RNA将通过与柱结合的RNA茎环结构的亲和力来选择。识别茎环的RNA物种(“适配子”)将从柱子上洗脱、逆转录、扩增和重新选择，以获得与茎环高亲和力的种群。将确定所选适配子(S)识别的结构域(表位)。这项研究的成功完成将使基于RNA的“免疫化学”的发展能够识别RNA中的同源或类似的三维结构，就像传统的抗体识别其他分子中的结构一样。这将是有价值的，因为RNA在体内是一个很差的抗原。%的RNA分子的部分功能是通过在溶液中呈现三维结构来实现的。为了开发分析RNA三维结构的新工具，将选择识别其他RNA三维结构的RNA分子。最终，RNA将被测试其识别两个RNA酶中结构相似结构域的能力，这两个RNA酶被假设具有相似的三维(三级)结构，尽管它们没有明显的一级序列相似性。***

9319139 Schmidt It has been shown recently that nucleic acid molecules can fold into a variety of shapes, capable of specific interaction with small molecule or protein ligands. This application proposes to synthesize a complex population of RNA species from a library of DNA templates containing approximately 10 (exp13) random sequences. The RNAs will be selected by affinity to a column-bound RNA stem- loop structure. RNA species recognizing the stem-loop ("aptamers") will be eluted from the column, reverse transcribed, amplified, and re-selected to obtain a population of high affinity to the stem- loop. The domain (epitope) recognized by selected aptamer(s) will be determined. The successful completion of this research will allow the development of an RNA-based "immunochemistry" capable of recognizing homologous or analogous three-dimensional structures in RNA, much as conventional antibodies recognize structures in other molecules. This would be valuable, since RNA is a poor antigen in vivo. %%% RNA molecules function in part by assuming three-dimensional structures in solution. In an effort to develop new tools for analyzing RNA three-dimensional structure, RNA molecules will be selected which recognize the three-dimensional structures of other RNAs. Eventually, RNAs will be tested for their ability to recognize structurally similar domains in two RNA enzymes which are hypothesized to have similar three-dimensional (tertiary) structures even though they do not have obvious primary sequence similarities. ***