Targeted Profiling of RNA Translation.

Curr Protoc Mol Biol
Authors
Keywords
Abstract

This unit describes a reverse transcription-quantitative PCR (RT-qPCR)-based method for gene-targeted measurement of RNA translation levels. The method includes washing and lysing cells with a buffer containing cycloheximide to enrich ribosomal accumulation at translation initiation sites (TIS), followed by enzymatic treatment to generate ribosomal footprints, reverse transcription targeted towards TIS of specific transcripts of interest to generate complementary DNA (cDNA), and qPCR to measure the abundance of these footprints. This method enables time- and cost-effective assessment of changes in translation levels across focused panels of genes and across numerous samples. © 2018 by John Wiley & Sons, Inc.

Year of Publication
2019
Journal
Curr Protoc Mol Biol
Volume
125
Issue
1
Pages
e71
Date Published
2019 Jan
ISSN
1934-3647
DOI
10.1002/cpmb.71
PubMed ID
30346115
PubMed Central ID
PMC6326074
Links
Grant list
R01 CA187918 / CA / NCI NIH HHS / United States
P50 CA168504 / CA / NCI NIH HHS / United States
R35 CA210057 / CA / NCI NIH HHS / United States
P50 CA165962 / CA / NCI NIH HHS / United States