An Integrated Polysome Profiling and Ribosome Profiling Method to Investigate In Vivo Translatome

Methods Mol Biol. 2018:1712:1-18. doi: 10.1007/978-1-4939-7514-3_1.

Abstract

Recent advances in global translatome analysis technologies enable us to understand how translational regulation of gene expression modulates cellular functions. In this chapter, we present an integrated method to measure various aspects of translatome by polysome profiling and ribosome profiling using purified B cells. We standardized our protocols to directly compare the results from these two approaches. Parallel assessment of translatome with these two approaches can generate a comprehensive picture on how translational regulation determines protein output.

Keywords: Polysome profiling; Ribo-seq; Ribosome profiling; Translational regulation of gene expression; Translatome.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • B-Lymphocytes / metabolism
  • Gene Expression Profiling / methods*
  • Models, Animal
  • Polyribosomes / genetics*
  • Protein Biosynthesis / genetics*
  • Proteins / genetics
  • RNA, Messenger / chemistry
  • RNA, Messenger / metabolism
  • Ribosomes / chemistry
  • Ribosomes / genetics*
  • Sequence Analysis, RNA

Substances

  • Proteins
  • RNA, Messenger