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mRNA Processing

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Cover of 'mRNA Processing'

Table of Contents

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    Book Overview
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    Chapter 1 Detection and Identification of Uncapped RNA by Ligation-Mediated Reverse Transcription Polymerase Chain Reaction
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    Chapter 2 Preparation of Splicing Competent Nuclear Extract from Mammalian Cells and In Vitro Pre-mRNA Splicing Assay
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    Chapter 3 Single-Molecule Analysis of Pre-mRNA Splicing with Colocalization Single-Molecule Spectroscopy (CoSMoS)
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    Chapter 4 In Vitro Modulation of Endogenous Alternative Splicing Using Splice-Switching Antisense Oligonucleotides
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    Chapter 5 Purification of mRNA Processing Complexes Using an RNA Affinity Approach
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    Chapter 6 Polyadenylation Site-Based Analysis of Transcript Expression by 3′READS+
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    Chapter 7 Comprehensive Identification of mRNA Polyadenylation Sites by PAPERCLIP
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    Chapter 8 Microinjection and Fluorescence In Situ Hybridization Assay for Studying mRNA Export in Mammalian Cells
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    Chapter 9 Methods for the Detection of Adenosine-to-Inosine Editing Events in Cellular RNA
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    Chapter 10 Using RNA-Seq to Discover Genetic Polymorphisms That Produce Hidden Splice Variants
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    Chapter 11 Inducible Expression of Eukaryotic Circular RNAs from Plasmids
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    Chapter 12 Methods for Extraction of RNA, Proteins, or Protein Complexes from Subcellular Compartments of Eukaryotic Cells
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    Chapter 13 Isolation of Newly Transcribed RNA Using the Metabolic Label 4-Thiouridine
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    Chapter 14 Robust, Cost-Effective Profiling of RNA Binding Protein Targets with Single-end Enhanced Crosslinking and Immunoprecipitation (seCLIP)
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    Chapter 15 Purification of Transcript-Specific mRNP Complexes Formed In Vivo from Saccharomyces cerevisiae
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    Chapter 16 A Cell-Based High-Throughput Method for Identifying Modulators of Alternative Splicing
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    Chapter 17 Genome-Wide RNAi Screens for RNA Processing Events in Drosophila melanogaster S2 Cells
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    Chapter 18 Assaying RNA Structure Inside Living Cells with SHAPE
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    Chapter 19 Erratum to: Robust, Cost-Effective Profiling of RNA Binding Protein Targets with Single-end Enhanced Crosslinking and Immunoprecipitation (seCLIP)
Attention for Chapter 5: Purification of mRNA Processing Complexes Using an RNA Affinity Approach
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Chapter title
Purification of mRNA Processing Complexes Using an RNA Affinity Approach
Chapter number 5
Book title
mRNA Processing
Published in
Methods in molecular biology, January 2017
DOI 10.1007/978-1-4939-7204-3_5
Pubmed ID
Book ISBNs
978-1-4939-7203-6, 978-1-4939-7204-3
Authors

Xiuye Wang, Yongsheng Shi

Abstract

Multiple mRNA processing steps, including splicing and 3' processing, take place in macromolecular complexes that contain many proteins and sometimes RNA molecules. A key challenge in the mRNA processing field has been to define the structure-function relationship of these sophisticated molecular machines. A prerequisite for addressing this challenge is to develop tools for purifying mRNA processing complexes in their native and intact forms that are suitable for functional and structural studies. Among many methods that have been developed, RNA affinity-based methods are most widely applied. In these methods, RNA molecules that are substrates to mRNA processing machineries are fused with an affinity tag, incubated with cellular extracts/lysates to allow for the assembly of mRNA processing complexes, and finally the assembled complexes are purified using RNA affinity tag. In this chapter, we will overview RNA affinity-based purification methods and describe in detail one such method, MS2-tagging, and its application in the purification of mRNA 3' processing complexes. Although these methods were originally developed for purifying mRNA processing complexes, they should be applicable to purification of other RNA-protein complexes as well.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 6 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 6 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 2 33%
Unspecified 1 17%
Student > Bachelor 1 17%
Other 1 17%
Unknown 1 17%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 2 33%
Unspecified 1 17%
Agricultural and Biological Sciences 1 17%
Engineering 1 17%
Unknown 1 17%