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      Image-Based Screening for Stress Granule Regulators.

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          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.

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          Author and article information

          Journal
          Journal ID (iso-abbrev): Methods Mol Biol
          Title: Methods in molecular biology (Clifton, N.J.)
          Publisher: Springer Science and Business Media LLC
          ISSN (Electronic): 1940-6029
          ISSN (Print): 1064-3745
          Publication date (Electronic): 2022
          Volume: 2428
          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.
          Article
          DOI: 10.1007/978-1-0716-1975-9_22
          PubMed ID: 35171491
          SO-VID: ad19292f-1edf-40a3-acc1-9ee0446c415c
          History

          Keywords: Z score,Microscopic screen,Automated image analysis,Stress granule,siRNA
          Data availability:
          Keywords: Z score, Microscopic screen, Automated image analysis, Stress granule, siRNA

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