Ribosome profiling: a tool for quantitative evaluation of dynamics in mRNA translation

Methods in Molecular Biology
Piyada JuntawongJulia Bailey-Serres

Abstract

Translational regulation is important for plant growth, metabolism, and acclimation to environmental challenges. Ribosome profiling involves the nuclease digestion of mRNAs associated with ribosomes and mapping of the generated ribosome-protected footprints to transcripts. This is useful for investigation of translational regulation. Here we present a detailed method to generate, purify, and high-throughput-sequence ribosome footprints from Arabidopsis thaliana using two different isolation methods, namely, conventional differential centrifugation and the translating ribosome affinity purification (TRAP) technology. These methodologies provide researchers with an opportunity to quantitatively assess with high-resolution the translational activity of individual mRNAs by determination of the position and number of ribosomes in the corresponding mRNA. The results can provide insights into the translation of upstream open reading frames, alternatively spliced transcripts, short open reading frames, and other aspects of translation.

Citations

Nov 1, 2017·Proceedings of the National Academy of Sciences of the United States of America·Jérémie BazinJulia Bailey-Serres
Jul 17, 2020·Frontiers in Plant Science·Federico Martinez-SeidelJoachim Kopka
Oct 12, 2019·Science·Mauricio A ReynosoJulia Bailey-Serres
Jun 13, 2018·Proceedings of the National Academy of Sciences of the United States of America·Elaine YeungRashmi Sasidharan
Apr 11, 2019·Frontiers in Plant Science·Takeshi FukaoJulián Mario Peña-Castro

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