Laser Targeted Oligo Ligation (LTOL) to Identify DNA Sequences in the Vicinity of a Single Subnuclear Structure in a Single Cell

Methods Mol Biol. 2020:2175:11-21. doi: 10.1007/978-1-0716-0763-3_2.

Abstract

Gene loci are organized around nuclear substructures, forming gene hubs which provide a level of transcriptional control. To date, most techniques used to investigate the genes in these hubs have been based on using material from bulk cells. This makes identifying specific gene associations difficult. Here we describe the Laser Targeted Oligo Ligation (LTOL) technique that was developed to identify DNA sequences around a single subnuclear structure on a single-cell basis by targeting these regions with two-photon irradiation.

Keywords: Gene hubs; Microscopy; Nuclear organization; Sequencing; Single cell; Subnuclear structures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence*
  • Cell Compartmentation
  • Cell Nucleus / genetics*
  • Cell Nucleus / metabolism*
  • Chromatin / metabolism
  • Gene Expression Regulation
  • Genetic Loci
  • HeLa Cells
  • Histones / genetics
  • Histones / metabolism
  • Humans
  • Immunohistochemistry / methods*
  • Lasers
  • Microscopy, Fluorescence / methods*
  • Oligodeoxyribonucleotides / genetics
  • Single-Cell Analysis / methods

Substances

  • Chromatin
  • Histones
  • Oligodeoxyribonucleotides
  • oligodeoxyadenylic acid

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