Ohuchi Shoji, Suess Beatrix
Department of Biology, Technische Universität Darmstadt, Germany.
FEBS Lett. 2017 May;591(10):1429-1436. doi: 10.1002/1873-3468.12653. Epub 2017 May 2.
RNA aptamers are one of the promising components for constructing artificial genetic circuits. In this study, we developed a transcriptional activator based on an RNA aptamer against one of the most frequently applied repressor proteins, lambda phage cI. In vitro selection (Systematic Evolution of Ligands by EXponential enrichment) and following in vivo screening identified an RNA aptamer with the intended transcriptional activator activity from an RNA pool containing a 40-nucleotide long random region. Quantitative analysis showed a 35-fold elevation of reporter expression upon aptamer expression. These results suggest that the diversity of artificial transcriptional activators can be extended by employing RNA aptamers against repressor proteins to broaden the parts for constructing genetic circuits.
RNA适配体是构建人工遗传回路的有前景的组件之一。在本研究中，我们基于一种针对最常用的阻遏蛋白之一——λ噬菌体cI的RNA适配体开发了一种转录激活剂。体外筛选（指数富集配体系统进化）及随后的体内筛选从一个包含40个核苷酸长随机区域的RNA文库中鉴定出了具有预期转录激活活性的RNA适配体。定量分析表明，适配体表达后报告基因的表达提高了35倍。这些结果表明，通过使用针对阻遏蛋白的RNA适配体可以扩展人工转录激活剂的多样性，从而拓宽构建遗传回路的元件。
