An Optimized Adaptation of DamID for NGS Applications

Methods Mol Biol. 2025:2866:245-262. doi: 10.1007/978-1-0716-4192-7_14.

Abstract

Recent studies have implicated higher-order genome organization in the regulation of genes and cellular state. Lamina-Associated Domains (LADs) are regions of heterochromatin associated with the nuclear envelope and the nuclear lamina, a protein network involved in both nuclear organization and genome structure. LADs are developmentally regulated, and their dysregulation is associated with several diseases and pathological states, including cancer and premature aging. In addition to LADs, other nuclear protein compartments appear to scaffold or support unique chromatin environments to affect gene expression. These revelations carry profound implications for our comprehension of developmental processes and the pathogenesis of various diseases, especially given the numerous disorders already directly associated with, for example, mutations in lamin and INM proteins. This spatial compartmentalization of chromatin subtypes to unique protein compartments has led to the adoption of proximity-labeling methods, such as DamID (DNA Adenine Methyltransferase Identification), to identify these unique chromatin compartments.

Keywords: Chromatin; DamID sequencing; Epigenetics; Genome Organization; Lamina-Associated Domains (LADs).

MeSH terms

  • Chromatin* / genetics
  • Chromatin* / metabolism
  • DNA Methylation
  • Heterochromatin / genetics
  • Heterochromatin / metabolism
  • High-Throughput Nucleotide Sequencing* / methods
  • Humans
  • Nuclear Lamina / genetics
  • Nuclear Lamina / metabolism
  • Site-Specific DNA-Methyltransferase (Adenine-Specific) / genetics
  • Site-Specific DNA-Methyltransferase (Adenine-Specific) / metabolism

Substances

  • Chromatin
  • Site-Specific DNA-Methyltransferase (Adenine-Specific)
  • Heterochromatin