2024-07-23 ワシントン大学セントルイス校
<関連情報>
- https://artsci.wustl.edu/ampersand/washu-chemists-develop-groundbreaking-test-track-crucial-edits-rna
- https://pubs.acs.org/doi/10.1021/acscentsci.4c00444
エンドヌクレアーゼV免疫染色アッセイ(EndoVIA)による細胞内のA-to-I編集の空間的可視化 Spatial Visualization of A-to-I Editing in Cells Using Endonuclease V Immunostaining Assay (EndoVIA)
Alexandria L. Quillin,Benoît Arnould,Steve D. Knutson,Jennifer M. Heemstra
ACS Central Science Published:July 7, 2024
DOI:https://doi.org/10.1021/acscentsci.4c00444
Abstract
Adenosine-to-inosine (A-to-I) editing is one of the most widespread post-transcriptional RNA modifications and is catalyzed by adenosine deaminases acting on RNA (ADARs). Varying across tissue types, A-to-I editing is essential for numerous biological functions, and dysregulation leads to autoimmune and neurological disorders, as well as cancer. Recent evidence has also revealed a link between RNA localization and A-to-I editing, yet understanding of the mechanisms underlying this relationship and its biological impact remains limited. Current methods rely primarily on in vitro characterization of extracted RNA that ultimately erases subcellular localization and cell-to-cell heterogeneity. To address these challenges, we have repurposed endonuclease V (EndoV), a magnesium-dependent ribonuclease that cleaves inosine bases in edited RNA, to selectively bind and detect A-to-I edited RNA in cells. The work herein introduces an endonuclease V immunostaining assay (EndoVIA), a workflow that provides spatial visualization of edited transcripts, enables rapid quantification of overall inosine abundance, and maps the landscape of A-to-I editing within the transcriptome at the nanoscopic level.
