生物技术通报 ›› 2023, Vol. 39 ›› Issue (4): 227-235.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0940

• 技术与方法 • 上一篇    下一篇

低起始量的免疫共沉淀技术研究进展

张新博(), 崔浩亮, 史佩华, 高锦春, 赵顺然, 陶晨雨()   

  1. 河北农业大学动物科技学院,保定 071000
  • 收稿日期:2022-07-28 出版日期:2023-04-26 发布日期:2023-05-16
  • 通讯作者: 陶晨雨,女,博士,副教授,研究方向:动物遗传育种与繁殖;E-mail: taochenyuty@163.com
  • 作者简介:张新博,女,硕士研究生,研究方向:动物遗传育种与繁殖;E-mail: zhangxinbo58@163.com
  • 基金资助:
    国家自然科学基金项目(32002175);河北省自然科学基金项目(C2020204058);河北农业大学引进人才科研专项(ZD201718)

Research Progress in Low-input Chromatin Immunoprecipitation Assay

ZHANG Xin-bo(), CUI Hao-liang, SHI Pei-hua, GAO Jin-chun, ZHAO Shun-ran, TAO Chen-yu()   

  1. College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000
  • Received:2022-07-28 Published:2023-04-26 Online:2023-05-16

摘要:

将染色质免疫共沉淀技术(chromatin immunoprecipitation assay, ChIP)和第二代测序技术相结合的染色体免疫共沉淀测序技术(co-immunoprecitation followed by sequencing, ChIP-seq)是分析全基因组表观遗传学变化的重要方法,它可以快速、有效地检测到在全基因组范围内DNA与蛋白结合位点、转录因子结合位点(transcription factor binding site, TFBS)、组蛋白翻译后修饰(histone post-translation modifications, hPTMs)、核小体定位和DNA甲基化等。然而,长期以来ChIP-seq需要大量细胞来生成高质量数据集,这限制了ChIP在一些细胞数较低的样本如卵母细胞、早期胚胎细胞等研究领域的应用。近年来,在ChIP的基础上,研究人员提出了一系列降低样品起始量和实验成本、提高测序质量的低起始量ChIP-seq方法,促进了表观基因组学的发展。本文综述了ChIP原理和降低起始量的ChIP的方法学发展,并对其中几种比较重要的方法进行了比较,并总结了低起始量的ChIP-seq在表观遗传学研究中的应用。本文最后对低起始量ChIP技术的应用和发展进行了展望,为低细胞数样品在低起始量ChIP的选择提供了参考。

关键词: 低起始量, 染色质免疫共沉淀, ChIP-seq, 转录因子, 组蛋白修饰

Abstract:

The chromosome immunoprecipitation sequencing technology(ChIP-seq), which combines the chromatin immunoprecipitation technology(ChIP)with the second generation sequencing technology,is an important method for analyzing the epigenetic changes of the whole genome,and can quickly and effectively detect DNA and protein binding sites,transcription factor binding sites(TFBS),histone post-translation modifications(hPTMs),nucleosome localization and DNA methylation in the whole genome. However,for a long time,ChIP-seq needs a large number of cells to generate high-quality data sets,which limits the application of ChIP in some specific tissue and low-cell samples such as oocytes,early embryonic cells and other research fields. In recent years, based on ChIP,researchers have proposed a series of low-input ChIP-seq methods to reduce the initial sample amount and experimental cost and increase the sequencing quality,which has promoted the development of epigenomics. In this paper, we reviewed the principles of ChIP and the methodological development of ChIP-seq reducing low-input amount,compared several of the more important methods,and summarized the application of ChIP-seq with low-input amount in epigenetic research. Finally,we prospected the application and development of low initial ChIP-seq technology,aiming to provide reference for the selection of different low-input ChIP-seq methods for low cell number samples.

Key words: low-input, chromatin co-immunoprecipitation, ChIP-seq, transcription factors, histone modifications