DNA甲基化测序技术研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2023-1166

摘要/Abstract

摘要：

DNA甲基化是一种重要的表观遗传修饰，在动植物生长发育、疾病发生、基因表达调控等方面发挥着关键作用。临床上，DNA甲基化肿瘤标志物可以作为肿瘤的诊断、预后和治疗的生物标志物。DNA甲基化的准确检测对于阐明生物生长发育、疾病发生等生命机理具有重要意义。DNA甲基化测序技术是研究DNA甲基化的有力工具，被广泛应用于DNA甲基化在基因组上的定位。近年来，为了更好地检测DNA甲基化位点信息，科学家提出了一系列DNA甲基化高通量测序技术方法，提高了测序检测灵敏度，降低了测序成本和实验花费，显著推动了表观基因组学的发展。本文综述了一系列DNA甲基化测序技术方法，重点介绍了WGBS、TAPS、EM-seq三种测序技术的技术原理及其应用场景，并介绍了在单细胞水平上定位DNA甲基化的测序方法，最后展望其未来发展的方向。

关键词: DNA甲基化, WGBS, TAPS, EM-seq, 单细胞, 多组学

Abstract:

DNA methylation is an important epigenetic modification that plays a key role in the growth and development of animals and plants, as well as in the occurrence of diseases and the regulation of gene expression. Clinically, DNA methylation tumor markers can serve as biological markers for the diagnosis, prognosis, and treatment of tumors. Accurate detection of DNA methylation is of great significance for elucidating the life mechanisms of biological growth, development, and disease occurrence. DNA methylation sequencing technology is a powerful tool for studying DNA methylation and is widely used for mapping DNA methylation in the genome. In recent years, in order to better detect DNA methylation site information, scientists have proposed a series of high-throughput DNA methylation sequencing technologies, which have improved sequencing detection sensitivity, reduced sequencing costs and experimental expenses, and greatly promoted the development of epigenomics. This article reviews a series of DNA methylation sequencing technologies, focusing on the technical principles and application scenarios of three sequencing technologies: WGBS, TAPS, and EM-seq. It also introduces sequencing methods for locating DNA methylation at the single-cell level, and finally prospects the future development direction.

Key words: DNA methylation, WGBS, TAPS, EM-seq, single cell, multiomics

袁明波, 叶国华, 杨丹, 宋冬雪. DNA甲基化测序技术研究进展[J]. 生物技术通报, 2024, 40(5): 58-65.

YUAN Ming-bo, YE Guo-hua, YANG Dan, SONG Dong-xue. Research Progress in DNA Methylation Sequencing Technology[J]. Biotechnology Bulletin, 2024, 40(5): 58-65.

图/表 5

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方法 Method	DNA片段化 DNA framentation	酶氧化 Enzymatic oxidation	强酸强碱Strong acids and alkalis	脱氨方式 Deamination method	扩增方式 Amplification method	文库丰富度 Library richness	数据分析软件 Data analysis software
WGBS	亚硫酸氢盐打断/超声打断	不使用	使用	亚硫酸氢盐	PCR	低	Bismark & BWAMeth
TAPS	超声打断	TET1氧化	不使用	吡啶硼烷	PCR	高	asTair
EM-seq	超声打断	TET2氧化	不使用	APOBEC3A脱氨酶	PCR	低	Bismark & BWAMeth

方法 Method	DNA片段化 DNA framentation	酶氧化 Enzymatic oxidation	强酸强碱Strong acids and alkalis	脱氨方式 Deamination method	扩增方式 Amplification method	文库丰富度 Library richness	数据分析软件 Data analysis software
WGBS	亚硫酸氢盐打断/超声打断	不使用	使用	亚硫酸氢盐	PCR	低	Bismark & BWAMeth
TAPS	超声打断	TET1氧化	不使用	吡啶硼烷	PCR	高	asTair
EM-seq	超声打断	TET2氧化	不使用	APOBEC3A脱氨酶	PCR	低	Bismark & BWAMeth