1.武汉大学 化学与分子科学学院,湖北 武汉 430072
2.武汉大学 公共卫生学院,湖北 武汉 430071
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陈梦园,游雪娇,袁必锋等.DNA甲基化修饰的定位分析方法研究进展[J].分析测试学报,2022,41(01):50-57.
CHEN Meng⁃yuan,YOU Xue⁃jiao,YUAN Bi⁃feng,et al.Analytical Methods for Localization of Methylated DNA Modification[J].Journal of Instrumental Analysis,2022,41(01):50-57.
陈梦园,游雪娇,袁必锋等.DNA甲基化修饰的定位分析方法研究进展[J].分析测试学报,2022,41(01):50-57. DOI: 10.19969/j.fxcsxb.21100501.
CHEN Meng⁃yuan,YOU Xue⁃jiao,YUAN Bi⁃feng,et al.Analytical Methods for Localization of Methylated DNA Modification[J].Journal of Instrumental Analysis,2022,41(01):50-57. DOI: 10.19969/j.fxcsxb.21100501.
5⁃甲基胞嘧啶(5⁃Methylcytosine,5mC)作为DNA中研究最为广泛的表观遗传修饰,不改变基因的序列,但是可以调控基因表达,从而在生命体的生长、发育和疾病发生中发挥着重要作用。研究5mC的分布、变化及作用机制有助于加强对生命体活动本质的理解。阐明基因组DNA中5mC修饰的生物学功能主要依赖于精准破译其在基因组上的位置信息。目前对5mC的定位分析除了经典的亚硫酸氢盐介导的方法之外,也发展了其他多种分析方法,如免疫沉淀富集介导的定位分析、酶介导的定位分析、吡啶硼烷介导的定位分析、纳米孔测序定位分析以及单分子实时测序定位分析等。该文总结了5mC定位分析方法的原理、优点和缺点,并对未来DNA甲基化修饰分析方法的发展方向进行了展望。
As the most widely studied epigenetic modification in DNA,5⁃methylcytosine(5mC) plays an important role in the growth and development of organisms.5mC participates the regulation of gene expressions without changing the sequences of genes.Investigation on the localization and content changes of 5mC in the incidence of diseases will strengthen our understanding toward its roles in the development of diseases.Elucidation of the biological functions of 5mC mainly depends on uncovering its accurate localization information in genomes.Over the past few decades,many analytical methods based on high⁃throughput sequencing technology have been established to localize 5mC in genomes.Apart from the classic BS⁃seq(bisulfite sequencing),some other analytical methods,such as DIP⁃seq(DNA immunoprecipitation sequencing),EM⁃seq(enzymatic methyl⁃seq),TAPS(TET⁃assisted pyridine borane sequencing),nanopore⁃seq(nanopore sequencing) and SMRT⁃seq(single molecule real⁃time) have been developed.In this review,the principles,advantages and disadvantages of these analytical methods for 5mC localization in DNA based on the high⁃throughput sequencing technology are summarized and discussed.In addition,the future research directions for mapping 5mC in DNA are also envisioned.It is hoped that this review will benefit and stimulate the study of the biological functions of 5mC in genomes.
DNA甲基化修饰5⁃甲基胞嘧啶定位分析测序
methylated DNA modification5⁃methylcytosinelocation analysissequencing
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