Using Chromatin Accessibility to Delineate Therapeutic Subtypes in Pancreatic Cancer Patient-Derived Cell Lines

STAR Protoc. 2020 Aug 4;1(2):100079. doi: 10.1016/j.xpro.2020.100079. eCollection 2020 Sep 18.

Abstract

Disrupted chromatin regulatory processes contribute to the development of cancer, in particular pancreatic ductal adenocarcinoma. The assay for transposase accessible chromatin with high-throughput sequencing (ATAC-seq) is typically used to study chromatin organization. Here, we present a revised ATAC-seq protocol to study chromatin accessibility in adherent patient-derived cell lines. We provide details on how to calculate the library molarity using Agilent's Bioanalyzer and an analysis pipeline for peak calling and transcription factor mapping. For complete details on the use and execution of this protocol, please refer to Brunton et al. (2020).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin / physiology*
  • Chromatin Immunoprecipitation Sequencing / methods*
  • Gene Expression / genetics
  • Gene Expression Regulation, Neoplastic / genetics
  • Gene Library
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Nucleosomes
  • Pancreas / metabolism
  • Pancreas / pathology
  • Pancreatic Neoplasms / classification
  • Pancreatic Neoplasms / physiopathology
  • Primary Cell Culture / methods*
  • Sequence Analysis, DNA / methods
  • Transcription Factors / metabolism
  • Transposases / metabolism

Substances

  • Chromatin
  • Nucleosomes
  • Transcription Factors
  • Transposases