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Chromatin Immunoprecipitation to Investigate H2A.Z Dynamics in Response to Environmental Changes
J Paola Saldierna Guzmán1, Mark Zander2, Björn C Willige3
1Department of Soil and Crop Sciences, College of Agricultural Sciences, Colorado State University, Fort Collins, CO, USA.
Abstract:
DNA methylation and posttranslational modifications of histones instruct gene expression in eukaryotes. Besides canonical histones, histone variants also play a critical role in transcriptional regulation. One of the best studied histone variants in plants is H2A.Z whose removal from gene bodies correlates with increased transcriptional activity. The eviction of H2A.Z is regulated by environmental cues such as increased ambient temperatures, and current models suggest that H2A.Z functions as a transcriptional buffer preventing environmentally responsive genes from undesired activation. To monitor temperature-dependent H2A.Z dynamics, chromatin immunoprecipitation (ChIP) of H2A.Z-occupied DNA can be performed. The following protocol describes a quick and easy ChIP approach to study in vivo H2A.Z occupancy.
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Chromatin Immunoprecipitation- ChIP
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ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...
Euchromatin
Euchromatin is the less dense region of the chromatin and stains lighter. Euchromatin contains histone H3 extensively...
Heterochromatin
Constitutive heterochromatin: It is a highly compact region of chromatin that is mostly concentrated in the centromere and telomere. Unlike euchromatin, the amino acid at...

