Use of Alu element containing minigenes to analyze circular RNAs

Justin R. Welden, Anna Pawluchin, Jacob van Doorn, Stefan Stamm

Research output: Contribution to journalArticlepeer-review

3 Scopus citations


In addition to linear mRNAs, many eukaryotic genes generate circular RNAs. Most circular RNAs are generated by joining a 5' splice site with an upstream 3' splice site within a pre-mRNA, a process called back-splicing. This circularization is likely aided by secondary structures in the pre-mRNA that bring the splice sites into close proximity. In human genes, Alu elements are thought to promote these secondary RNA structures, as Alu elements are abundant and exhibit base complementarities with each other when present in opposite directions in the pre-mRNA. Here, we describe the generation and analysis of large, Alu element containing reporter genes that form circular RNAs. Through optimization of cloning protocols, reporter genes with up to 20 kb insert length can be generated. Their analysis in co-transfection experiments allows the identification of regulatory factors. Thus, this method can identify RNA sequences and cellular components involved in circular RNA formation.

Original languageEnglish
Article numbere59760
JournalJournal of Visualized Experiments
Issue number157
StatePublished - Mar 2020

Bibliographical note

Publisher Copyright:
© 2020 Journal of Visualized Experiments.


  • Alternative splicing
  • Biochemistry
  • Circular RNAs
  • Issue 157
  • MRNA
  • Minigene
  • RNA
  • Reporter gene

ASJC Scopus subject areas

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


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