Analysis of Chromatin Accessibility, Histone Modifications, and Transcriptional States in Specific Cell Types Using Flow Cytometry

Methods Mol Biol. 2023:2698:57-73. doi: 10.1007/978-1-0716-3354-0_5.

Abstract

The past two decades in biomedical research have experienced an explosion of cell type-specific and single-cell studies, especially concerning the concomitant dissection of regulatory and transcriptional landscapes of those under investigation. Additionally, leveraging next-generation sequencing (NGS) platforms efforts have been undertaken to evaluate the effects of chromatin accessibility, histone modifications, or even transcription factor binding sites. We have shown that Fluorescence-Activated Nuclear Sorting (FANS) is an effective means to characterize the transcriptomes of nuclei from different tissues. In light of our own technical and experimental developments, we extend this effort to combine FACS/FANS with Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq), Chromatin Immunoprecipitation sequencing (ChIP-seq), and RNA sequencing (RNA-seq) for profiling individual cell types according to their chromatin and transcriptional states.

Keywords: ATAC-seq; ChIP-seq; FACS; FANS; NGS; Nuclei; Protoplasts; RNA-seq.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus
  • Chromatin* / genetics
  • Flow Cytometry
  • Histone Code*
  • Protein Processing, Post-Translational

Substances

  • Chromatin