An Ex Vivo Protocol to Assess IRE1*and符号*alpha;-Dependent RNA Cleavage Using Total RNA Isolated from Mouse Tissues
Year: 2025
Type: article
Abstract: . Quantitative assessment is achieved using agarose gel electrophoresis and imaging software. This methodology enables the study of IRE1α's RNA cleavage activity under conditions that closely mimic in vivo environments, providing a more physiologically relevant approach to understanding the role of RIDD in cellular and tissue-specific contexts. Key features • Uses total RNA from mouse tissues instead of synthetic RNA to better reflect in vivo conditions. • Includes RIDD-specific controls such as IRE1α inhibitor (4μ8C) and RNase A to confirm targeted RNA cleavage. • Combines agarose gel electrophoresis and ImageJ quantification for both qualitative and statistical validation. • Allows comparative studies of IRE1α activity across multiple mouse tissues in different biological contexts.
Source: BIO-PROTOCOL
Authors: Mohammad Rashid, So-Young Rah, Se-Woong Ko, Han-Jung Chae
Institution: Jeonbuk National University
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: diamond
OpenAlex ID: W4412590853