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Updated: Dec 6, 2025
10:37
Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
Published on: November 15, 2017
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In vivo mRNA display enables large-scale proteomics by next generation sequencing
Panos Oikonomou1,2,3, Roberto Salatino2, Saeed Tavazoie1,2,3
1Department of Biological Sciences, Columbia University, New York, NY 10027; po2194@columbia.edu st2744@columbia.edu.
Summary
This study introduces in vivo mRNA display, a novel proteomics technology. It uses DNA sequencing to efficiently identify proteins and their functions in their native cellular environment.
Area of Science:
- Molecular Biology
- Proteomics
- Biotechnology
Background:
- Proteomics requires scalable, standardized, and cost-effective methods for functional protein characterization in native cellular contexts.
- Current proteomics approaches, particularly mass spectrometry, face limitations in scalability and cost compared to genomics.
- There is a need for advanced technologies to bridge the gap between genomic and proteomic capabilities.
Purpose of the Study:
- To introduce and validate a novel technology, in vivo mRNA display, for large-scale proteomic analysis.
- To demonstrate the capability of in vivo mRNA display for high-throughput protein identification and functional characterization.
- To showcase the potential of this technology in systematically characterizing native functional proteomes.
Main Methods:
- Development of in vivo mRNA display technology coupling expressed proteins with their encoding messenger RNAs (mRNAs).
- Utilizing a high-affinity stem-loop RNA binding domain interaction for mRNA-protein complex formation.
- Leveraging next-generation DNA sequencing for high-throughput, sensitive, and specific protein identification.
Main Results:
- Generation of a high-coverage in vivo mRNA display library for the Saccharomyces cerevisiae proteome.
- Demonstration of the technology's effectiveness in characterizing subcellular localization of proteins.
- Successful application in identifying protein interactions within their native cellular context.
Conclusions:
- In vivo mRNA display transforms proteomics applications into DNA sequencing problems, enhancing scalability and reducing costs.
- This technology overcomes key limitations of mass spectrometry-based proteomics.
- It offers a powerful platform for systematically characterizing native functional proteomes by leveraging advances in DNA sequencing.
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