<script type="text/javascript">
<!--
document.write('<div id="oa_widget"></div>');
document.write('<script type="text/javascript" src="https://www.openaire.eu/index.php?option=com_openaire&view=widget&format=raw&projectId=undefined&type=result"></script>');
-->
</script>

COPY SCRIPT

For further information contact us at helpdesk@openaire.eu

In vitro RNA Cleavage Assays to Characterize IRE1-dependent RNA Decay

descriptionPublicationkeyboard_double_arrow_right Article , Other literature type 01 Jan 2019Publisher:Bio-Protocol, LLCJournal:BIO-PROTOCOL, volume 9 (issn: 2331-8325,

Authors: Diego Acosta-Alvear; Jirka Peschek; Jirka Peschek; Peter Walter; Peter Walter; G. Elif Karagöz;

doi: 10.21769/bioprotoc.3307

pmid: 33654817

pmc: PMC7854161

In vitro RNA Cleavage Assays to Characterize IRE1-dependent RNA Decay

- Summary
- Subjects
- Metrics

Abstract

The kinase/RNase IRE1 is a key effector of the cellular response to endoplasmic reticulum stress. The RNase activity of IRE1 can be measured in cells or in the test tube. Here we describe a protocol for the in vitro cleavage and analysis of RNA substrates of IRE1. The method consists of the in vitro transcription, purification and re-folding of IRE1 substrate RNAs followed by their cleavage using recombinant cytosolic kinase/RNase domains of IRE1 and the separation of the resulting fragments by denaturing polyacrylamide gel electrophoresis. This protocol allows the study of the cleavage kinetics of IRE1's RNA substrates in vitro.

Related Organizations

University of California System
United States
Medical University of Vienna
Austria
University of California, Santa Barbara
United States
University of California San Francisco Medical Center
United States
University of California, San Francisco
United States

View all View all

Keywords

QH301-705.5, Methods Article, Biology (General)

Impact byBIP!

	citations This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).	2
	popularity This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.	Average
	influence This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).	Average
	impulse This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.	Average