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Turbo-RIP: A Protocol for TurboID-based RNA Immunopurification to Map RNA Landscapes in Plant Biomolecular
Zhi Zhang1, Yanting Xu1, Hanxiang Liu1
1State Key Laboratory of Biocontrol, Guangdong Key Laboratory of Plant Stress Biology, Innovation center for evolutionary synthetic biology, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China.
Bio-Protocol
|February 12, 2026
Summary
Researchers developed Turbo-RIP, a new method to identify RNAs within plant biomolecular condensates. This technique helps understand plant stress responses by capturing transiently associated RNAs.
Area of Science:
- Molecular Biology
- Plant Science
- Biochemistry
Background:
- Biomolecular condensates are essential for cellular processes, organizing proteins and RNAs via liquid-liquid phase separation.
- Identifying RNA composition within these condensates is crucial for understanding plant stress responses.
- Current methods for capturing transiently associated RNAs in plant condensates are technically challenging.
Purpose of the Study:
- To present Turbo-RIP (TurboID-based proximity labeling with RNA immunopurification), a comprehensive protocol for identifying condensate-associated RNAs in plants.
- To provide detailed procedures for implementing Turbo-RIP in various plant species and experimental conditions.
Main Methods:
- Turbo-RIP utilizes TurboID biotin ligase for proximity labeling of proteins at 22 °C.
- Formaldehyde crosslinking and streptavidin-based capture of protein-RNA complexes are employed.
- The protocol includes procedures for cloning strategies, plant transformation (Nicotiana benthamiana, Arabidopsis thaliana), TurboID validation, and RNA recovery.
Main Results:
- The Turbo-RIP protocol successfully captured processing body-associated RNAs with minimal background noise.
- The method is adaptable for different plant species and condensate types.
- Quality control measures and data analysis pipelines are integrated into the protocol.
Conclusions:
- Turbo-RIP offers a robust and accessible method for systematic mapping of RNA populations within plant condensates.
- This protocol facilitates the study of plant stress responses and other cellular processes regulated by biomolecular condensates.
- The 3-5 day protocol is suitable for standard plant molecular biology laboratories.
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