Highly quantitative measurement of differential protein-genome binding with PerCell chromatin sequencing.

Cell Rep Methods

Nationwide Children's Hospital, Center for Childhood Cancer Research, Columbus, OH, USA; Department of Pediatrics, The Ohio State University College of Medicine, Columbus, OH, USA; Department of Biological Chemistry & Pharmacology, The Ohio State University College of Medicine, Columbus, OH, USA. El

Published: June 2025


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Article Abstract

Quantitative comparison of ChIP-seq profiling between experimental conditions or samples remains technically challenging for the epigenetics field. Here, we report a strategy combining the use of well-defined cellular spike-in ratios of orthologous species' chromatin and a bioinformatic analysis pipeline to facilitate highly quantitative comparisons of 2D chromatin sequencing across experimental conditions. We find that the PerCell methodology results in efficient and consistent levels of spike-in vs. experimental genomic reads. We demonstrate use of the method and pipeline to enable quantitative, internally normalized chromatin sequencing on zebrafish embryos and human cancer cells. Overall, we propose the PerCell method to enable cross-species comparative epigenomics and promote uniformity of data analyses and sharing across labs.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC12272247PMC
http://dx.doi.org/10.1016/j.crmeth.2025.101052DOI Listing

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