RNA Aptamers Recognizing RNA Structural Elements

小行星9319139施密特 最近已经显示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特异性相互作用。 本申请提出从含有大约10个（exp 13）随机序列的DNA模板文库合成RNA种类的复杂群体。 通过对柱结合的RNA茎环结构的亲和力来选择RNA。 识别茎-环的RNA种类（“适体”）将从柱洗脱、逆转录、扩增和重新选择以获得对茎-环具有高亲和力的群体。 将确定由所选适体识别的结构域（表位）。 这项研究的成功完成将允许开发一种基于RNA的“免疫化学”，能够识别RNA中的同源或类似三维结构，就像传统抗体识别其他分子中的结构一样。 这将是有价值的，因为RNA在体内是一种不良抗原。 % RNA分子在溶液中部分通过呈现三维结构来发挥作用。 为了开发用于分析RNA三维结构的新工具，将选择识别其他RNA的三维结构的RNA分子。 最终，RNA将被测试其识别两种RNA酶中结构相似结构域的能力，这两种RNA酶被假设具有相似的三维（三级）结构，即使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. ***