Institut de Chimie Moléculaire, ICMUB CNRS UMR6302, UBFC Dijon, France.
Faculty of Pharmaceutical Sciences, The University of British Columbia, Vancouver, Canada.
Nucleic Acids Res. 2019 Jun 20;47(11):5502-5510. doi: 10.1093/nar/gkz215.
Guanine-rich DNA and RNA sequences can fold into higher-order structures known as G-quadruplexes (or G4-DNA and G4-RNA, respectively). The prevalence of the G4 landscapes in the human genome, transcriptome and ncRNAome (non-coding RNA), collectively known as G4ome, is strongly suggestive of biological relevance at multiple levels (gene expression, replication). Small-molecules can be used to track G4s in living cells for the functional characterization of G4s in both normal and disease-associated changes in cell biology. Here, we describe biotinylated biomimetic ligands referred to as BioTASQ and their use as molecular tools that allow for isolating G4s through affinity pull-down protocols. We demonstrate the general applicability of the method by purifying biologically relevant G4s from nucleic acid mixtures in vitro and from human cells through the G4RP-RT-qPCR protocol. Overall, the results presented here represent a step towards the optimization of G4-RNAs identification, a key step in studying G4s in cell biology and human diseases.
富含鸟嘌呤的 DNA 和 RNA 序列可以折叠成称为 G-四链体（或分别为 G4-DNA 和 G4-RNA）的高级结构。在人类基因组、转录组和 ncRNA 组（非编码 RNA）中，G4 景观的普遍性强烈表明在多个层面上具有生物学相关性（基因表达、复制）。小分子可用于在活细胞中追踪 G4，以对 G4 在细胞生物学的正常和与疾病相关的变化中的功能特征进行功能表征。在这里，我们描述了称为 BioTASQ 的生物模拟配体，并将其用作分子工具，通过亲和下拉协议分离 G4。我们通过 G4RP-RT-qPCR 方案从体外核酸混合物和人类细胞中纯化具有生物学意义的 G4，证明了该方法的普遍适用性。总的来说，这里呈现的结果代表了朝着优化 G4-RNAs 鉴定迈出的一步，这是研究细胞生物学和人类疾病中 G4 的关键步骤。
