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具有营养应用的DNA甲基化分析技术。

Techniques of DNA methylation analysis with nutritional applications.

作者信息

Mansego Maria L, Milagro Fermín I, Campión Javier, Martínez J Alfredo

机构信息

CIBERobn, Physiopathology of Obesity and Nutrition, Institute of Health Carlos III, Madrid, Spain.

出版信息

J Nutrigenet Nutrigenomics. 2013;6(2):83-96. doi: 10.1159/000350749. Epub 2013 May 8.

Abstract

Epigenetic mechanisms are likely to play an important role in the regulation of metabolism and body weight through gene-nutrient interactions. This review focuses on methods for analyzing one of the most important epigenetic mechanisms, DNA methylation, from single nucleotide to global measurement depending on the study goal and scope. In addition, this study highlights the major principles and methods for DNA methylation analysis with emphasis on nutritional applications. Recent developments concerning epigenetic technologies are showing promising results of DNA methylation levels at a single-base resolution and provide the ability to differentiate between 5-methylcytosine and other nucleotide modifications such as 5-hydroxymethylcytosine. A large number of methods can be used for the analysis of DNA methylation such as pyrosequencing™, primer extension or real-time PCR methods, and genome-wide DNA methylation profile from microarray or sequencing-based methods. Researchers should conduct a preliminary analysis focused on the type of validation and information provided by each technique in order to select the best method fitting for their nutritional research interests.

摘要

表观遗传机制可能通过基因-营养相互作用在代谢和体重调节中发挥重要作用。本综述重点介绍了根据研究目标和范围,从单核苷酸到全局测量来分析最重要的表观遗传机制之一——DNA甲基化的方法。此外,本研究强调了DNA甲基化分析的主要原理和方法,并重点介绍了其在营养领域的应用。表观遗传技术的最新进展显示了在单碱基分辨率下DNA甲基化水平的有前景的结果,并提供了区分5-甲基胞嘧啶和其他核苷酸修饰(如5-羟甲基胞嘧啶)的能力。大量方法可用于DNA甲基化分析,如焦磷酸测序™、引物延伸或实时PCR方法,以及基于微阵列或测序的全基因组DNA甲基化图谱分析方法。研究人员应进行初步分析,重点关注每种技术提供的验证类型和信息,以便选择最适合其营养研究兴趣的最佳方法。

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