β-珠蛋白基因远程相互作用元件的筛选与功能性分析

Screening and functional analysis of the long-range interaction elements of β-globin genes

目的:研究表明β-珠蛋白基因在发育过程中呈现选择表达的转换机制,其上游的基因座位点控制区(LCR)调控了β-珠蛋白基因家族的表达模式。为了深入探究β-珠蛋白基因表达调控的分子网络,本文对其他可能参与调控β-珠蛋白基因表达的远程调控元件进行筛选,对β-珠蛋白基因动态调控转换机制进行深入研究。方法:早幼粒细胞经全反式维甲酸诱导分化,以β-珠蛋白基因启动子区及LCR作为环形染色体构象捕获(4C)技术分析的靶位点,通过测序技术与调控元件分析结合,在全基因组筛选与β-珠蛋白家族基因座位发生相互作用关系的位点。结果:根据4C测序结果,筛选出与HBD启动子区及LCR存在相互作用关系的位点。通过染色体构象捕获(3C)验证,结果与测序结果一致。通过甲醛辅助分离调控元件技术及Epiregio在线网站对调控元件功能性分析,结果表明筛选位点AC105129.4、AL354707.17、AC078785.22及AC021646.35均为潜在的参与β-珠蛋白基因调控元件。结论:4C筛选位点与锚定位点相互作用表明了β-珠蛋白家族基因座位在细胞核内复杂的空间组织形式。

Objective:Studies have shown that β-globin gene presents a selective expression transformation mechanism during development,and its upstream locus control region(LCR)regulates the expression pattern of β-globin gene family. To further explore the molecular network of β-globin gene expression regulation,other long-range regulatory elements that may be involved in the regulation of β-globin gene expression were screened and the dynamic regulation and transformation mechanism of β-globin gene was deeply studied. Methods:Promyelocytic cells were induced to differentiate by all-trans retinoic acid. β-globin gene promoter region and LCR were used as the target sites for circular chromosome conformational capture(4C) analysis.Through sequencing and regulatory element analysis,the sites interacting with β-globin family loci were screened in the whole genome. Results:According to the results of 4C sequencing,the sites that interact with HBD promoter region and LCR were screened. Verified by chromosome conformational capture(3C),the results were consistent with those of sequencing. The functional analysis of regulatory elements by formaldehyde-assisted separation regulatory elements and Epiregio online website showed that the screening sites AC105129.4,AL354707.17,AC078785.22 and AC021646.35 were all potential regulatory elements involved in β-globin gene. Conclusion:The interaction between 4C screening site and anchor site showed the complex spatial organization of β-globin family loci in the nucleus.