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Single and Combined Methods to Specifically or Bulk-Purify RNA-Protein Complexes
Roosje Van Ende1, Sam Balzarini1, Koen Geuten1
1Molecular Biotechnology of Plants and Micro-organisms, KU Leuven, Kasteelpark Arenberg 31, 3001 Leuven, Belgium.
Biomolecules
|August 14, 2020
Summary
This review explores RNA- and ribonucleoprotein-(RNP)-centric methods for studying the ribonome. It discusses strategies to cross-link RNA to proteins and overcome challenges in RNP yield and purity.
Area of Science:
- Molecular Biology
- Biochemistry
- Genomics
Background:
- The ribonome links the proteome and transcriptome, hosting crucial biological processes.
- Studying the ribonome can be done via omics or by targeting individual RNA or protein species.
Purpose of the Study:
- To review RNA- and ribonucleoprotein-(RNP)-centric methods for ribonome analysis.
- To discuss strategies for cross-linking RNA to proteins and studying RNP dynamics.
- To identify and propose solutions for challenges in RNP yield, purity, and cost.
Main Methods:
- Focus on RNA-centric and RNP-centric methodologies.
- Exploration of strategies to cross-link RNA to proteins.
- Utilizing organic phase separation protocols like XRNAX, OOPS, and PTex.
Main Results:
- Challenges in RNP yield, purity, and experimental cost are common across methods.
- Combining existing approaches can overcome these difficulties.
- Novel organic phase separation protocols offer solutions.
Conclusions:
- Effective study of the ribonome requires robust RNA-protein cross-linking and RNP analysis methods.
- Integration of advanced separation techniques is key to improving RNP studies.
- Future research should focus on optimizing these combined approaches for broader application.
