哺乳动物细胞DNA高级结构G-四链体检测方法的建立

The Establishment of the Detection Method of G-Quadruplex of DNA Structure in Mammalian Cells

G-四链体(G-quadruplex,G4)结构是由富含鸟嘌呤(G)的寡核苷酸折叠堆积形成的四链螺旋结构,在维持基因组稳定性和调控基因表达上有重要意义。然而,目前细胞内检测G-四链体方法有限,该文利用G-四链体特异性抗体(BG4)对经细胞质抽提和RNase处理的贴壁培养细胞基因组G-四链体进行免疫荧光–荧光原位杂交检测,直接观察DNA高级结构G-四链体,实现了细胞水平G-四链体的可视化,这将有助于今后更准确、直观地在细胞水平研究DNA G-四链体,明确G-四链体与小分子配体相互作用,巩固G-四链体在遗传学、表观遗传学和病理学等领域研究中的地位。

Guanine-rich oligonucleotides folded into quadruple-stranded helical structures known as G-quadruplexs（G4）, which have been associated with genomic stability and the regulation of gene expression. However, to date, the methods of imaging G-quadruplexes in mammalian cells remain poorly defined. Here, we reported the generation and application of a structure-specific antibody（BG4） that was employed to visualize DNA G-quadruplex structures and telomeric G-quadruplex by immunofluorescence-fluorescence in situ hybridization（IF-FISH） in adherent cells with extracting and treating by RNase. This is contributing to research of G-quadruplex structure more accurately and intuitively in vivo, definituding the interaction of between G-quadruplex（telomeric G-quadruplex） and ligands, and consolidating the position of G-quadruplex structure in the areas of genetics, epigenetics and pathology.