利用染色质免疫沉淀技术分析急性髓系白血病中的组蛋白修饰

Analysis of Histone Modifications in Acute Myeloid Leukaemia Using Chromatin Immunoprecipitation.

作者信息

Shields Benjamin J, Keniry Andrew, Blewitt Marnie E, McCormack Matthew P

机构信息

Australian Centre for Blood Diseases, Monash University, Melbourne, VIC, Australia.

The Walter and Eliza Hall Institute, Parkville, VIC, Australia.

出版信息

Methods Mol Biol. 2018;1725:177-184. doi: 10.1007/978-1-4939-7568-6_15.

DOI:10.1007/978-1-4939-7568-6_15

PMID:29322418

Abstract

Chromatin Immunoprecipitation (ChIP) using antibodies specific for histone modifications is a powerful technique for assessing the epigenetic states of cell populations by either quantitative PCR (ChIP-PCR) or next generation sequencing analysis (ChIP-Seq). Here we describe the procedure for ChIP of histone marks in myeloid leukaemia cell lines and the subsequent purification of genomic DNA associated with repressive and activating histone modifications for further analysis. This procedure can be widely applied to a variety of histone marks to assess both activating and repressive modifications in the context of myeloid leukaemia.

摘要

使用针对组蛋白修饰的特异性抗体进行染色质免疫沉淀（ChIP），是一种通过定量PCR（ChIP-PCR）或下一代测序分析（ChIP-Seq）来评估细胞群体表观遗传状态的强大技术。在此，我们描述了在髓系白血病细胞系中进行组蛋白标记ChIP的程序，以及随后纯化与抑制性和激活性组蛋白修饰相关的基因组DNA以进行进一步分析的方法。该程序可广泛应用于多种组蛋白标记，以评估髓系白血病背景下的激活性和抑制性修饰。