Precise Antibody-Independent m6A Identification via 4SedTTP-Involved and FTO-Assisted Strategy at Single-Nucleotide Resolution

J Am Chem Soc. 2018 May 9;140(18):5886-5889. doi: 10.1021/jacs.7b13633. Epub 2018 Mar 5.

Abstract

Innovative detection techniques to achieve precise m6A distribution within mammalian transcriptome can advance our understanding of its biological functions. We specifically introduced the atom-specific replacement of oxygen with progressively larger atoms (sulfur and selenium) at 4-position of deoxythymidine triphosphate to weaken its ability to base pair with m6A, while maintaining A-T* base pair virtually the same as the natural one. 4SedTTP turned out to be an outstanding candidate that endowed m6A with a specific signature of RT truncation, thereby making this "RT-silent" modification detectable with the assistance of m6A demethylase FTO through next-generation sequencing. This antibody-independent, 4SedTTP-involved and FTO-assisted strategy is applicable in m6A identification, even for two closely gathered m6A sites, within an unknown region at single-nucleotide resolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / chemistry*
  • Antibodies / metabolism
  • DNA, Single-Stranded / chemistry*
  • DNA, Single-Stranded / metabolism
  • Humans
  • Methyltransferases / analysis*
  • Methyltransferases / metabolism
  • Selenium / chemistry*
  • Selenium / metabolism
  • Thymine Nucleotides / chemistry*
  • Thymine Nucleotides / metabolism

Substances

  • Antibodies
  • DNA, Single-Stranded
  • Thymine Nucleotides
  • Methyltransferases
  • METTL3 protein, human
  • Selenium
  • thymidine 5'-triphosphate