Protocol for base resolution mapping of ac4C using RedaC:T-seq

David Sturgill*, Daniel Arango*, Shalini Oberdoerffer*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

N4-acetylcytidine (ac4C) is an mRNA modification catalyzed by the enzyme N-acetyltransferase 10 (NAT10), with position-dependent effects on mRNA translation. This protocol details a procedure to map ac4C at base resolution using NaBH4-induced reduction of ac4C and conversion to thymidine followed by sequencing (RedaC:T-seq). Total RNA is ribodepleted and then treated with NaBH4 to reduce ac4C to tetrahydro-ac4C, which specifically alters base pairing during cDNA synthesis, allowing the detection of ac4C at positions called as thymidine following Illumina sequencing. For complete details on the use and execution of this protocol, please refer to Arango et al. (2022).1

Original languageEnglish (US)
Article number101858
JournalSTAR Protocols
Volume3
Issue number4
DOIs
StatePublished - Dec 16 2022

Keywords

  • Bioinformatics
  • Molecular Biology
  • RNAseq
  • Sequencing

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology
  • General Neuroscience

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