期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 60, 期 31, 页码 17009-17017出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202102762
关键词
adenosine-to-inosine editing; bioorganic chemistry; chemiluminescence; immunoassays; RNA
资金
- National Institutes of Health [R21GM134564, R01GM140657]
- Emory Integrated Genomics Core (EIGC) - Emory University School of Medicine
The study presents an EndoV-linked immunosorbency assay (EndoVLISA) for rapid measurement of global A-to-I editing signatures in cellular RNA. The method is cost-effective, straightforward, and suitable for disease detection and drug development applications.
Adenosine-to-inosine (A-to-I) editing is a conserved eukaryotic RNAmodification 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.
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