Department of Chemistry and Konstanz Research School Chemical Biology (KoRS-CB), University of Konstanz, 78457 Konstanz, Germany.
Methods. 2012 May;57(1):115-21. doi: 10.1016/j.ymeth.2012.02.005. Epub 2012 Feb 23.
DNA and RNA G-quadruplexes have gained increasing attention due to their potential role in a wide range of biological functions. The majority of functional studies characterize the influence of quadruplexes in gene expression including transcription and translation. Many of these studies have used reporter assays to elucidate the effect of quadruplexes at certain positions in promoters and untranslated mRNA regions (UTRs). Reporter assays are the preferred method to ascertain the biological function of DNA or RNA G-quadruplexes intracellularly due to their ready availability, fast cloning and experimental setup and reproducibility. Moreover, these reporter assays are also helpful to compare or screen for selectivity and efficacy of small molecules that target DNA and RNA G-quadruplexes in the cellular context. Here we briefly discuss various aspects of reporter assays followed by a review of available studies using reporter assays to understand the role and functions of DNA and RNA quadruplexes in gene expression.
DNA 和 RNA 四链体由于其在广泛的生物学功能中的潜在作用而受到越来越多的关注。大多数功能研究都描述了四链体对基因表达的影响,包括转录和翻译。这些研究中的许多都使用报告基因测定来阐明四链体在启动子和未翻译 mRNA 区域 (UTRs) 中某些位置的影响。由于报告基因测定具有易于获得、快速克隆和实验设置以及可重复性等优点,因此它们是确定 DNA 或 RNA G-四链体在细胞内的生物学功能的首选方法。此外,这些报告基因测定还有助于在细胞环境中比较或筛选针对 DNA 和 RNA G-四链体的小分子的选择性和功效。在这里,我们简要讨论了报告基因测定的各个方面,然后回顾了使用报告基因测定来了解 DNA 和 RNA 四链体在基因表达中的作用和功能的现有研究。