Image-Based Screening for Stress Granule Regulators

Katharina Hoerth; Nina Eiermann; Jürgen Beneke; Holger Erfle; Georg Stoecklin

doi:10.1007/978-1-0716-1975-9_22

Image-Based Screening for Stress Granule Regulators

Methods Mol Biol. 2022:2428:361-379. doi: 10.1007/978-1-0716-1975-9_22.

Authors

Katharina Hoerth^#^{1

2}, Nina Eiermann^#^{1

2}, Jürgen Beneke^{3

4}, Holger Erfle^{3

4}, Georg Stoecklin^{5

6

7}

Affiliations

¹ Division of Biochemistry, Medical Faculty Mannheim, Mannheim Institute for Innate Immunoscience (MI3), Heidelberg University, Mannheim, Germany.
² Center for Molecular Biology of Heidelberg University (ZMBH), German Cancer Research Center (DKFZ)-ZMBH Alliance, Heidelberg, Germany.
³ Advanced Biological Screening Facility, BioQuant, Heidelberg University, Heidelberg, Germany.
⁴ CellNetworks Cluster of Excellence, Heidelberg University, Heidelberg, Germany.
⁵ Division of Biochemistry, Medical Faculty Mannheim, Mannheim Institute for Innate Immunoscience (MI3), Heidelberg University, Mannheim, Germany. Georg.Stoecklin@medma.uni-heidelberg.de.
⁶ Center for Molecular Biology of Heidelberg University (ZMBH), German Cancer Research Center (DKFZ)-ZMBH Alliance, Heidelberg, Germany. Georg.Stoecklin@medma.uni-heidelberg.de.
⁷ CellNetworks Cluster of Excellence, Heidelberg University, Heidelberg, Germany. Georg.Stoecklin@medma.uni-heidelberg.de.

^# Contributed equally.

PMID: 35171491
DOI: 10.1007/978-1-0716-1975-9_22

Abstract

Stress granule (SG)-based RNA interference (RNAi) screening is a powerful method to discover factors that control protein synthesis and aggregation, as well as regulators of SG assembly and disassembly. Here, we describe how to set up and optimize a large-scale siRNA screen, and give a detailed outline for the automated quantification of SGs as a visual readout. Hit evaluation via calculated Z scores provides a list of candidates for further in-depth studies.

Keywords: Automated image analysis; Microscopic screen; Stress granule; Z score; siRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cytoplasmic Granules* / metabolism
Protein Biosynthesis
RNA Interference
RNA, Small Interfering / genetics
RNA, Small Interfering / metabolism
Stress Granules*
Stress, Physiological

Substances

RNA, Small Interfering