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Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation

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Cover of 'Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation'

Table of Contents

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    Book Overview
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    Chapter 1 Fluorescence Reporter-Based Genome-Wide RNA Interference Screening to Identify Alternative Splicing Regulators.
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    Chapter 2 Tandem Affinity Purification Approach Coupled to Mass Spectrometry to Identify Post-translational Modifications of Histones Associated with Chromatin-Binding Proteins.
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    Chapter 3 Efficient Preparation of High-Complexity ChIP-Seq Profiles from Early Xenopus Embryos.
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    Chapter 4 Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation
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    Chapter 5 Assay for Transposase-Accessible Chromatin with High-Throughput Sequencing (ATAC-Seq) Protocol for Zebrafish Embryos.
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    Chapter 6 Establishment of Time- and Cell-Specific RNAi in Caenorhabditis elegans.
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    Chapter 7 Cell-Penetrating Peptide-Mediated Delivery of Cas9 Protein and Guide RNA for Genome Editing.
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    Chapter 8 Epigenetic Analysis of Endocrine Cell Subtypes from Human Pancreatic Islets.
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    Chapter 9 eIF3 Regulation of Protein Synthesis, Tumorigenesis, and Therapeutic Response.
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    Chapter 10 High-Resolution Gene Expression Profiling of RNA Synthesis, Processing, and Decay by Metabolic Labeling of Newly Transcribed RNA Using 4-Thiouridine.
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    Chapter 11 Accurate Detection of Differential Expression and Splicing Using Low-Level Features.
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    Chapter 12 Profiling Changes in Histone Post-translational Modifications by Top-Down Mass Spectrometry.
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    Chapter 13 Determining if an mRNA is a Substrate of Nonsense-Mediated mRNA Decay in Saccharomyces cerevisiae.
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    Chapter 14 Optimizing In Vitro Pre-mRNA 3' Cleavage Efficiency: Reconstitution from Anion-Exchange Separated HeLa Cleavage Factors and from Adherent HeLa Cell Nuclear Extract.
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    Chapter 15 Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation
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    Chapter 16 Using an Inducible CRISPR-dCas9-KRAB Effector System to Dissect Transcriptional Regulation in Human Embryonic Stem Cells.
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    Chapter 17 In Vitro Assay to Study Histone Ubiquitination During Transcriptional Regulation.
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    Chapter 18 Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation
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    Chapter 19 Large-Scale RNA Interference Screening to Identify Transcriptional Regulators of a Tumor Suppressor Gene.
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    Chapter 20 Transcriptional Analysis-Based Integrative Genomics Approach to Identify Tumor-Promoting Metabolic Genes.
Attention for Chapter 14: Optimizing In Vitro Pre-mRNA 3' Cleavage Efficiency: Reconstitution from Anion-Exchange Separated HeLa Cleavage Factors and from Adherent HeLa Cell Nuclear Extract.
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Chapter title
Optimizing In Vitro Pre-mRNA 3' Cleavage Efficiency: Reconstitution from Anion-Exchange Separated HeLa Cleavage Factors and from Adherent HeLa Cell Nuclear Extract.
Chapter number 14
Book title
Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation
Published in
Methods in molecular biology, January 2017
DOI 10.1007/978-1-4939-6518-2_14
Pubmed ID
27832541
Book ISBNs
978-1-4939-6516-8, 978-1-4939-6518-2
Authors

Mihwa Na, Susana T. Valente, Kevin Ryan

Editors

Narendra Wajapeyee, Romi Gupta

Abstract

Eukaryotic RNA processing steps during mRNA maturation present the cell with opportunities for gene expression regulation. One such step is the pre-mRNA 3' cleavage reaction, which defines the downstream end of the 3' untranslated region and, in nearly all mRNA, prepares the message for addition of the poly(A) tail. The in vitro reconstitution of 3' cleavage provides an experimental means to investigate the roles of the various multi-subunit cleavage factors. Anion-exchange chromatography is the simplest procedure for separating the core mammalian cleavage factors. Here we describe a method for optimizing the in vitro reconstitution of 3' cleavage activity from the DEAE-sepharose separated HeLa cleavage factors and show how to ensure, or avoid, dependence on creatine phosphate. Important reaction components needed for optimal processing are discussed. We also provide an optimized procedure for preparing small-scale HeLa nuclear extracts from adherent cells for use in 3' cleavage in vitro.

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Mendeley readers

The data shown below were compiled from readership statistics for 3 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 3 100%

Demographic breakdown

Readers by professional status Count As %
Professor 1 33%
Student > Ph. D. Student 1 33%
Student > Doctoral Student 1 33%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 2 67%
Pharmacology, Toxicology and Pharmaceutical Science 1 33%

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