Trans ligation of RNAs to generate hybrid circular RNAs using highly efficient autocatalytic transcripts. Academic Article uri icon

Overview

abstract

  • Circular RNAs are useful entities for various biotechnology applications, such as templating translation and binding or sequestering miRNA and RNA binding proteins. Circular RNA as highly resistant to degradation in cells and are more long-lived than linear RNAs. Here, we describe a method for intracellular trans ligation of RNA transcripts that can generate hybrid circular RNAs. These hybrid circular RNAs comprise two separate RNA that are covalently linked by ligation to form a circular RNA. By incorporating self-cleaving ribozymes at each site of ligation, trans ligation of the transcripts occurs in mammalian cells with no additional material. We provide a protocol for designing and testing trans ligation of transcripts and demonstrate detection of hybrid circular RNAs using fluorescence microscopy.

publication date

  • May 13, 2021

Research

keywords

  • RNA, Catalytic
  • RNA, Circular

Identity

PubMed Central ID

  • PMC8589881

Scopus Document Identifier

  • 85107280367

Digital Object Identifier (DOI)

  • 10.1093/nar/gkl151

PubMed ID

  • 33992775

Additional Document Info

volume

  • 196