Preparation of Circular RNA In Vitro

Methods Mol Biol. 2018:1724:181-192. doi: 10.1007/978-1-4939-7562-4_15.

Abstract

This chapter describes a simple and straightforward way to obtain single-stranded circular RNA sequences in vitro. Linear RNA that is phosphorylated at the 5' end is first prepared by a chemical or enzymatic method, then circularized using ligase. The function of the prepared circular RNA molecule, such as an ability to induce translation, can then be investigated.

Keywords: Circular RNA; In vitro transcription; Ligase; Phosphorylation; Polyacrylamide gel electrophoresis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Ligases / metabolism*
  • Exoribonucleases / metabolism*
  • Humans
  • In Vitro Techniques
  • RNA / biosynthesis*
  • RNA / chemistry
  • RNA / genetics*
  • RNA Ligase (ATP) / metabolism*
  • RNA, Circular

Substances

  • RNA, Circular
  • RNA
  • Exoribonucleases
  • DNA Ligases
  • RNA Ligase (ATP)