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Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling
Published on: October 28, 2014
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Polysome profiling followed by quantitative PCR for identifying potential micropeptide encoding long non-coding RNAs
1MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, Guangdong 510275, China.
STAR Protocols
|January 3, 2022
Summary
Researchers developed a cost-effective method using polysome profiling to identify micropeptide-encoding long non-coding RNAs (lncRNAs). This technique aids in discovering novel regulators of cellular processes, advancing molecular biology research.
Area of Science:
- Molecular Biology
- Genomics
- Cellular Biology
Background:
- Micropeptides regulate diverse cellular functions.
- Long non-coding RNAs (lncRNAs) can encode micropeptides via small reading frames.
- Current detection methods for translating lncRNAs are complex and costly.
Purpose of the Study:
- To present an accessible and economical protocol for screening micropeptide-encoding lncRNAs.
- To enable simultaneous identification of multiple translating lncRNAs.
Main Methods:
- Utilized polysome profiling in suspension cell lines.
- Combined polysome profiling with quantitative PCR (qPCR).
Main Results:
- Developed an easy and cost-effective screening protocol.
- Facilitated simultaneous identification of numerous translating lncRNAs.
Conclusions:
- The presented protocol offers a practical approach for identifying micropeptide-encoding lncRNAs.
- This method can advance the study of micropeptides and their regulatory roles in cellular processes.
