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Measuring Translation Efficiency by RNA Immunoprecipitation of Translation Initiation Factors

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Cell Cycle Checkpoints

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2267))

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Abstract

Eukaryotic mRNAs are bound by a multitude of RNA binding proteins (RBPs) that control their localization, transport, and translation. Measuring the rate of translation of mRNAs is critical for understanding the factors and pathways involved in gene expression. In this chapter, we present a method to compare the rate of translation of individual mRNA species based on the fraction of mRNA bound by a specific ribonucleoprotein involved in the general translation machinery. The ribonucleoprotein complex is immunoprecipitated using an antibody for the RBP, followed by RT-PCR to semi-quantitatively determine the amount of an individual mRNA fraction bound by a translation regulating protein such as eIF4E.

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Author notes

  1. Chris Lucchesi and Shakur Mohibi contributed equally to this work.

Authors and Affiliations

  1. Comparative Oncology Laboratory, University of California at Davis, Davis, CA, USA

    Chris Lucchesi, Shakur Mohibi & Xinbin Chen

Authors
  1. Chris Lucchesi
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  2. Shakur Mohibi
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  3. Xinbin Chen
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Corresponding author

Correspondence to Xinbin Chen .

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Editors and Affiliations

  1. Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    James J. Manfredi

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Lucchesi, C., Mohibi, S., Chen, X. (2021). Measuring Translation Efficiency by RNA Immunoprecipitation of Translation Initiation Factors. In: Manfredi, J.J. (eds) Cell Cycle Checkpoints. Methods in Molecular Biology, vol 2267. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1217-0_5

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  • DOI: https://doi.org/10.1007/978-1-0716-1217-0_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1216-3

  • Online ISBN: 978-1-0716-1217-0

  • eBook Packages: Springer Protocols

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