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Peroxisomes

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Cover of 'Peroxisomes'

Table of Contents

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    Book Overview
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    Chapter 1 Isolation of Peroxisomes from Rat Liver and Cultured Hepatoma Cells by Density Gradient Centrifugation
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    Chapter 2 Isolation of Peroxisomes from Mouse Brain Using a Continuous Nycodenz Gradient: A Comparison to the Isolation of Liver and Kidney Peroxisomes
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    Chapter 3 Determining the Topology of Peroxisomal Proteins Using Protease Protection Assays
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    Chapter 4 Isolation of Native Soluble and Membrane-Bound Protein Complexes from Yeast Saccharomyces cerevisiae.
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    Chapter 5 Method for Measurement of Peroxisomal Very Long-Chain Fatty Acid Beta-Oxidation and De Novo C26:0 Synthesis Activity in Living Cells Using Stable-Isotope Labeled Docosanoic Acid
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    Chapter 6 Analysis of Plasmalogen Synthesis in Cultured Cells
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    Chapter 7 Transfection of Primary Human Skin Fibroblasts for Peroxisomal Studies
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    Chapter 8 siRNA-mediated Silencing of Peroxisomal Genes in Mammalian Cells
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    Chapter 9 Dual Reporter Systems for the Analysis of Translational Readthrough in Mammals
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    Chapter 10 Cytochemical Detection of Peroxisomes in Light and Electron Microscopy with 3,3′-diaminobenzidine
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    Chapter 11 Cryo-Immuno Electron Microscopy of Peroxisomal Marker Proteins
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    Chapter 12 Detection and Immunolabeling of Peroxisomal Proteins
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    Chapter 13 Labeling of Peroxisomes for Live Cell Imaging in the Filamentous Fungus Ustilago maydis.
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    Chapter 14 Quantitative Monitoring of Subcellular Redox Dynamics in Living Mammalian Cells Using RoGFP2-Based Probes
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    Chapter 15 KillerRed as a Tool to Study the Cellular Responses to Peroxisome-Derived Oxidative Stress
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    Chapter 16 Determination of Peroxisomal pH in Living Mammalian Cells Using pHRed
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    Chapter 17 In Cellulo Approaches to Study Peroxisomal Protein Import - Yeast Immunofluorescence Microscopy.
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    Chapter 18 Blue Native PAGE: Applications to Study Peroxisome Biogenesis
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    Chapter 19 In Vitro PMP Import Analysis Using Cell-Free Synthesized PMP and Isolated Peroxisomes
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    Chapter 20 Peroxisomal Membrane and Matrix Protein Import Using a Semi-Intact Mammalian Cell System
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    Chapter 21 The Use of Glycosylation Tags as Reporters for Protein Entry into the Endoplasmic Reticulum in Yeast and Mammalian Cells.
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    Chapter 22 Detection of Ubiquitinated Peroxisomal Proteins in Yeast.
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    Chapter 23 Assessing Pexophagy in Mammalian Cells.
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    Chapter 24 Experimental Systems to Study Yeast Pexophagy.
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    Chapter 25 Flow Cytometric Analysis of the Expression Pattern of Peroxisomal Proteins, Abcd1, Abcd2, and Abcd3 in BV-2 Murine Microglial Cells
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    Chapter 26 Study of Peroxisomal Protein Phosphorylation by Functional Proteomics.
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    Chapter 27 Analysis of Peroxisomal β-Oxidation During Storage Oil Mobilization in Arabidopsis thaliana Seedlings
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    Chapter 28 Peroxisome Mini-Libraries: Systematic Approaches to Study Peroxisomes Made Easy.
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    Chapter 29 Generation of Peroxisome-Deficient Somatic Animal Cell Mutants
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    Chapter 30 Clinical and Laboratory Diagnosis of Peroxisomal Disorders.
Attention for Chapter 8: siRNA-mediated Silencing of Peroxisomal Genes in Mammalian Cells
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Chapter title
siRNA-mediated Silencing of Peroxisomal Genes in Mammalian Cells
Chapter number 8
Book title
Peroxisomes
Published in
Methods in molecular biology, April 2017
DOI 10.1007/978-1-4939-6937-1_8
Pubmed ID
Book ISBNs
978-1-4939-6935-7, 978-1-4939-6937-1
Authors

Schrader, Tina A., Schrader, Michael, Tina A. Schrader, Michael Schrader

Editors

Michael Schrader

Abstract

RNAi technologies are a valuable tool in the identification and investigation of proteins that are involved in peroxisome biogenesis and function. Small interfering RNA (siRNA) has developed into the most commonly used RNAi tool for the induction of transient, short-term silencing of protein coding genes. Although siRNA can induce gene knockdown in a variety of mammalian cell lines, their utility is limited by efficient uptake of synthetic oligonucleotides into the cells. Here, we describe different transfection methods that have been successfully used by us to silence peroxisomal genes in a variety of cell lines, including primary human skin fibroblasts, which are usually difficult to transfect.

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Mendeley readers

The data shown below were compiled from readership statistics for 15 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 15 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 4 27%
Student > Master 3 20%
Student > Bachelor 2 13%
Unspecified 1 7%
Student > Doctoral Student 1 7%
Other 1 7%
Unknown 3 20%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 6 40%
Chemistry 3 20%
Agricultural and Biological Sciences 2 13%
Pharmacology, Toxicology and Pharmaceutical Science 1 7%
Unspecified 1 7%
Other 0 0%
Unknown 2 13%