Single-Cell 5fC Sequencing

Methods Mol Biol. 2019:1979:251-267. doi: 10.1007/978-1-4939-9240-9_16.

Abstract

Active DNA demethylation plays important roles in the epigenetic reprogramming of developmental processes. 5-formylcytosine (5fC) is produced during active demethylation of 5-methylcytosine (5mC). Here, we describe a technique called CLEVER-seq (Chemical-labeling-enabled C-to-T conversion sequencing), which detects the whole genome 5fC distribution at single-base and single-cell resolution. CLEVER-seq is suitable for the analysis of precious samples such as early embryos and laser microdissection captured samples.

Keywords: 5-Formylcytosine; Bisulfite-free sequencing; CLEVER-seq; Chemical labeling; Single cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cytosine / analogs & derivatives*
  • Cytosine / analysis
  • DNA / chemistry*
  • DNA / genetics*
  • DNA Methylation*
  • Gene Library
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Sequence Analysis, DNA / methods*
  • Single-Cell Analysis / methods*

Substances

  • 5-formylcytosine
  • Cytosine
  • DNA