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Capturing Chromatin Organization by MNase-seq and ATAC-seq.

Hox genes play a pivotal role during development. Their expression is tightly controlled in a spatiotemporal manner, ensuring that specific body structures develop at the correct locations and times during development. Various genomics approaches have been used to capture temporal and dynamic regulation of Hox gene expression at the nucleosome/chromatin level. This chapter focuses on the utilization of capture MNase-seq and Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq), two advanced techniques that enable the exploration of chromatin accessibility and nucleosome positioning within these critical genomic regions.

Chromatin

MNase-seq to Identify Genome-Wide DNA-Protein Interactions.

Identification of the occupancy of transcription factors (TFs) and nucleosomes across the genome yields insights into the regulation of gene expression patterns. While several independent techniques can be performed and then analyzed in composite to reveal this chromatin landscape, the use of micrococcal nuclease (MNase) digestion can resolve the footprints of nearly all chromatin proteins simultaneously. The protocol below describes the use of MNase to identify chromatin footprints of both TFs and nucleosomes in two vastly different cell types, Mouse embryonic stem cells (mESCs) and sperm, with differing levels of chromatin compaction.

Animals