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Title: Chromatin immunoprecipitation improvements for the processing of small frozen pieces of adipose tissue.
Authors: Castellano-Castillo, Daniel
Denechaud, Pierre-Damien
Moreno-Indias, Isabel
Tinahones, Francisco
Fajas, Lluis
Queipo-Ortuño, María Isabel
Cardona, Fernando
metadata.dc.subject.mesh: Adipose Tissue
Chromatin Immunoprecipitation
Genome, Human
Middle Aged
Real-Time Polymerase Chain Reaction
Issue Date: 14-Feb-2018
Abstract: Chromatin immunoprecipitation (ChIP) has gained importance to identify links between the genome and the proteome. Adipose tissue has emerged as an active tissue, which secretes a wide range of molecules that have been related to metabolic and obesity-related disorders, such as diabetes, cardiovascular failure, metabolic syndrome, or cancer. In turn, epigenetics has raised the importance in discerning the possible relationship between metabolic disorders, lifestyle and environment. However, ChIP application in human adipose tissue is limited by several factors, such as sample size, frozen sample availability, high lipid content and cellular composition of the tissue. Here, we optimize the standard protocol of ChIP for small pieces of frozen human adipose tissue. In addition, we test ChIP for the histone mark H3K4m3, which is related to active promoters, and validate the performance of the ChIP by analyzing gene promoters for factors usually studied in adipose tissue using qPCR. Our improvements result in a higher performance in chromatin shearing and DNA recovery of adipocytes from the tissue, which may be useful for ChIP-qPCR or ChIP-seq analysis.
metadata.dc.identifier.doi: 10.1371/journal.pone.0192314
Appears in Collections:Producción 2020

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