title: Measuring mRNA Translation by Polysome Profiling.
abstract: Determination of mRNA translation rates is essential to understanding the regulatory pathways governing eukaryotic gene expression. In this chapter, we present a transcriptome-wide method to assess translation by association of mRNAs with polysomes on sucrose density gradients. After sedimentation, the fractions are spiked with a control RNA mixture and the RNA content is measured by high-throughput sequencing. Normalization to the spike-ins provides a global quantitative view on the translational status of cellular mRNAs, with the ability to measure changes and identify active and silent subpopulations of each.
authors.affiliation: Department of Cell Biology and Neuroscience, University of California, Riverside, 900 University Avenue, Riverside, CA, 92521, USA.
authors.forename: Marek
authors.initials: M
authors.lastname: Kudla
authors.affiliation: Department of Cell Biology and Neuroscience, University of California, Riverside, 900 University Avenue, Riverside, CA, 92521, USA. karginov@ucr.edu.
authors.forename: Fedor V
authors.initials: FV
authors.lastname: Karginov
mesh_terms: D002499:Centrifugation, Density Gradient / Q000379:methods; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D011132:Polyribosomes / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D017423:Sequence Analysis, RNA / Q000379:methods; D059467:Transcriptome
chemical_list: D012333:RNA, Messenger
keywords: Polysome; Sucrose gradient; Translation; mRNA