Development of high throughput screening method of G-quadruplexes using small molecules

g -四plex （G4s）是在富含鸟嘌呤的多核苷酸中形成的三维结构。到目前为止，相对较少的g4形成寡核苷酸（GFOs）被发现，但它们在基因表达和复制的调控中起着重要作用。计算分析表明，基因组中存在大量的gfo。然而，目前还没有一种简单直接的方法来大规模筛选gfo。在本项目中，我们利用荧光G4配体L1Cy5-7OTD与DNA微阵列开发了一种高通量筛选GFOs的方法。通过这种方法，我们在CpG岛（CGI）微阵列中发现了1,998个新的gfo，其中包含88,737个靶标。基因本体（Gene Ontology， GO）分析表明，这些gfo主要与代谢过程、转录调控、生物合成过程和发育过程有关。该方法可以很容易地应用于其他DNA微阵列。

G-Quadruplexes (G4s) are three-dimensional structures that are formed in guanine-rich polynucleotides. Relatively few G4-forming oligonucleotides (GFOs) have been identified so far, but they are important because they play significant roles in regulation of gene expression and replication. Computational analysis suggests huge numbers of GFOs are present in the genome. However, there has been no simple and direct screening methodology for large-scale screening of GFOs.In this project, we have developed a high-throughput screening method for GFOs by using the fluorescent G4 ligand L1Cy5-7OTD with a DNA microarray. With this method, we found 1,998 novel GFOs in a CpG island (CGI) microarray bearing 88,737 targets. Gene Ontology (GO) analysis suggests that these GFOs were mainly associate with metabolic processes, transcriptional regulation, biosynthetic processes and developmental processes. The present method would be easily applicable to other DNA microarrays.