Translating Ribosome Affinity Purification (TRAP) followed by RNA sequencing technology (TRAP-SEQ) for quantitative assessment of plant translatomes

Methods in Molecular Biology
Mauricio A ReynosoM Eugenia Zanetti

Abstract

Translating Ribosome Affinity Purification (TRAP) is a technology to isolate the population of mRNAs associated with at least one 80S ribosome, referred as the translatome. TRAP is based on the expression of an epitope-tagged version of a ribosomal protein and the affinity purification of ribosomes and associated mRNAs using antibodies conjugated to agarose beads. Quantitative assessment of the translatome is achieved by direct RNA sequencing (RNA-SEQ), which provides accurate quantitation of ribosome-associated mRNAs and reveals alternatively spliced isoforms. Here we present a detailed procedure for TRAP, as well as a guide for preparation of RNA-SEQ libraries (TRAP-SEQ) and a primary data analysis. This methodology enables the study of translational dynamic by assessing rapid changes in translatomes, at organ or cell-type level, during development or in response to endogenous or exogenous stimuli.

Citations

Jun 27, 2017·Methods : a Companion to Methods in Enzymology·Xicotencatl Gracida, John A Calarco
Mar 16, 2018·Acta Pharmacologica Sinica·Xiao-Fen RuanYaoliang Tang
Aug 9, 2018·BMC Genomics·K BligheC B T Moore
May 27, 2020·Annual Review of Virology·Madhu KappagantuJames N Culver
Sep 19, 2017·Frontiers in Plant Science·Gaurav SablokKemal Kazan
May 6, 2020·Wiley Interdisciplinary Reviews. RNA·Ricardo A Urquidi CamachoAlbrecht G von Arnim
Apr 11, 2019·Frontiers in Plant Science·Takeshi FukaoJulián Mario Peña-Castro
Oct 12, 2019·Science·Mauricio A ReynosoJulia Bailey-Serres
Nov 20, 2016·Plant, Cell & Environment·Albrecht G von Arnim, Hanjo A Hellmann
Oct 17, 2017·Frontiers in Plant Science·David C HaakSong Li
Apr 20, 2021·The Plant Cell·Titouan Bonnot, Dawn H Nagel
Jul 24, 2021·The Biochemical Journal·Soledad TraubenikMaría Eugenia Zanetti
Jul 2, 2021·Plant, Cell & Environment·Mariana A S Artur, Kaisa Kajala

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