Direct Immunodetection of Global A-to-I RNA Editing Activity with a Chemiluminescent Bioassay

Steve D. Knutson, Robert A. Arthur, H. Richard Johnston, Jennifer M. Heemstra

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


Adenosine-to-inosine (A-to-I) editing is a conserved eukaryotic RNA modification that contributes to development, immune response, and overall cellular function. Here, we utilize Endonuclease V (EndoV), which binds specifically to inosine in RNA, to develop an EndoV-linked immunosorbency assay (EndoVLISA) as a rapid, plate-based chemiluminescent method for measuring global A-to-I editing signatures in cellular RNA. We first optimize and validate our assay with chemically synthesized oligonucleotides. We then demonstrate rapid detection of inosine content in treated cell lines, demonstrating equivalent performance against current standard RNA-seq approaches. Lastly, we deploy our EndoVLISA for profiling differential A-to-I RNA editing signatures in normal and diseased human tissue, illustrating the utility of our platform as a diagnostic bioassay. Together, the EndoVLISA method is cost-effective, straightforward, and utilizes common laboratory equipment, offering a highly accessible new approach for studying A-to-I editing. Moreover, the multi-well plate format makes this the first assay amenable for direct high-throughput quantification of A-to-I editing for applications in disease detection and drug development.

Original languageEnglish
Pages (from-to)17009-17017
Number of pages9
JournalAngewandte Chemie - International Edition
Issue number31
StatePublished - Jul 26 2021


  • RNA
  • adenosine-to-inosine editing
  • bioorganic chemistry
  • chemiluminescence
  • immunoassays


Dive into the research topics of 'Direct Immunodetection of Global A-to-I RNA Editing Activity with a Chemiluminescent Bioassay'. Together they form a unique fingerprint.

Cite this