An optimized ChIP-Seq framework for profiling of histone modifications in Chromochloris zofingiensis

التفاصيل البيبلوغرافية
العنوان: An optimized ChIP-Seq framework for profiling of histone modifications in Chromochloris zofingiensis
المؤلفون: Stefan Schmollinger, Ronan C. O'Malley, Matthew Mingay, Sabeeha S. Merchant, Cindy Chen, Daniela Strenkert
بيانات النشر: Cold Spring Harbor Laboratory, 2021.
سنة النشر: 2021
مصطلحات موضوعية: chemistry.chemical_compound, Histone, chemistry, biology.protein, H3K4me3, Epigenetics, Computational biology, Biology, Genome, Chromatin immunoprecipitation, Gene, DNA, Deep sequencing
الوصف: The eukaryotic green alga Chromochloris zofingiensis is a reference organism for studying carbon partitioning and a promising candidate for the production of biofuel precursors. Recent transcriptome profiling transformed our understanding of its biology and generally algal biology, but epigenetic regulation remains understudied and represents a fundamental gap in our understanding of algal gene expression. Chromatin Immunoprecipitation followed by deep sequencing (ChIP-Seq) is a powerful tool for the discovery of such mechanisms, by identifying genome-wide histone modification patterns and transcription factor-binding sites alike. Here, we established a ChIP-Seq framework for Chr. zofingiensis yielding over 20 million high quality reads per sample. The most critical steps in a ChIP experiment were optimized, including DNA shearing to obtain an average DNA fragment size of 250 bp and assessment of the recommended formaldehyde concentration for optimal DNA-protein crosslinking. We used this ChIP-Seq framework to generate a genome-wide map of the H3K4me3 distribution pattern and to integrate these data with matching RNA-Seq data. In line with observations from other organisms, H3K4me3 marks predominantly transcription start sites of genes. Our H3K4me3 ChIP-Seq data will pave the way for improved genome structural annotation in the emerging reference alga Chr. zofingiensis.
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::91bae573d4cef39e1558855a9018d391
https://doi.org/10.1101/2021.11.11.468265
حقوق: OPEN
رقم الأكسشن: edsair.doi...........91bae573d4cef39e1558855a9018d391
قاعدة البيانات: OpenAIRE