Direct Immunodetection of Global A-to-I RNA Editing Activity with a Chemiluminescent Bioassay.
Angew Chem Int Ed Engl
; 60(31): 17009-17017, 2021 07 26.
Article
em En
| MEDLINE
| ID: mdl-33979483
ABSTRACT
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 Endonucleaseâ
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.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
RNA
/
Adenosina
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Inosina
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Medições Luminescentes
Limite:
Humans
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article