Work

Targeted Profiling of RNA Translation

Year: 2018

Type: article

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.

Source: Current Protocols in Molecular Biology

DOI: 10.1002/cpmb.71

Authors: Ben B. Li, Changli Qian, Thomas M. Roberts, Jean J. Zhao

Institutions: Broad Institute, Harvard University, Dana-Farber Cancer Institute

Language: English

Topic: RNA and protein synthesis mechanisms

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: Breast Cancer Research Foundation, National Institutes of Health, National Cancer Institute