Quantification of MRNA Ribosomal Engagement in Human Neurons Using Parallel Translating Ribosome Affinity Purification (TRAP) and RNA Sequencing

Overview

Journal STAR Protoc

Publisher Cell Press

Specialties Biology
Biomedical Engineering
Science

Date 2020 Dec 28

PMID 33364619

Citations 3

Authors

Deivid Carvalho Rodrigues

Marat Mufteev

James Ellis

Affiliations

Soon will be listed here.

Abstract

Translation regulation is a fundamental step in gene regulation with critical roles in neurodevelopment. Here, we describe three protocols to calculate the ribosomal-engagement levels of the transcriptome from -derived neuronal cells. The protocols described here include enrichment of -generated pluripotent-derived neurons, immunoaffinity purification of ribosome-bound RNAs, and calculation of the fraction of ribosome-engaged mRNAs. The ribosome-engaged RNA fraction is a measurement of the translation activity, and differences between genotype or growth conditions report change in translational regulation. For complete details on the use and execution of this protocol, please refer to Rodrigues et al. (2020).

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