Studying the Translatome with Polysome Profiling
Abstract
Polysome fractionation by sucrose density gradient centrifugation followed by analysis of RNA and protein is a technique that allows to understand the changes in translation of individual mRNAs as well as genome-wide effects on the translatome. Here, we describe the polysome profiling technique and RNA as well as protein isolation procedures from sucrose fractions.
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Polysome profiling overcomes the limitation of single-variant-per-cell MPRAs by enabling single cells to translate tens to hundreds of in vitro transcribed (IVT) and transfected mRNA variants.
“Polysome profiling13 overcomes this limitation by enabling single cells to translate tens to hundreds of in vitro transcribed (IVT) and transfected mRNA variants.”
From Human 5′ UTR design and variant effect prediction from a massively parallel translation assay (2019) · cites this paper as [13]
Cites
Global quantification of mammalian gene expression control
Schwanhäusser, B., Busse, D., Li, N., Dittmar, G., Schuchhardt, J., Wolf, J., Chen, W., & Selbach, M. (2011) · Nature · doi:10.1038/nature10098 · Cited by 6,566 · see citing works
Kraushar, M. L., Thompson, K., Wijeratne, H. R. S., Viljetic, B., Sakers, K., Marson, J. W., Kontoyiannis, D. L., Buyske, S., Hart, R. P., & Rasin, M.-R. (2014) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1408305111 · Cited by 98 · see citing works
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Human 5′ UTR design and variant effect prediction from a massively parallel translation assay
Sample, P. J., Wang, B., Reid, D. W., Presnyak, V., McFadyen, I. J., Morris, D. R., & Seelig, G. (2019) · Nature Biotechnology · doi:10.1038/s41587-019-0164-5 · Cited by 442 · see citing works
- Authors
- Zuccotti, P., & Modelska, A.
- Affiliations
- University of Trento (IT)
- Corresponding author
- Modelska, Angelika
- ORCID
- Modelska, Angelika
- Venue
- Methods in Molecular Biology
- Publisher
- Springer New York
- Published
- 2016
- Volume
- 1358
- Pages
- 59-69
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- OpenAlex
- W2462718056
- Cited by
- 20
- References
- 2
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 1 · 2023: 2 · 2022: 2 · 2021: 3 · 2020: 4 · 2019: 1 · 2018: 2 · 2017: 1 · 2016: 1
- FWCI
- 2.13
- Type
- book chapter
- Language
- en
- Open access
- closed
- Licence
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
- Keywords
- polysome profiling, translatome, translation (biology), mRNA translation, sucrose density gradient centrifugation, polysome fractionation, RNA extraction, protein extraction, ribosome profiling
- MeSH terms
- Humans, Molecular Biology, Polyribosomes, Proteins, RNA, Messenger, Protein Biosynthesis, Genome
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