Download e-book for iPad: Alternative pre-mRNA Splicing: Theory and Protocols by Stefan Stamm, Chris Smith, Reinhard Lührmann

By Stefan Stamm, Chris Smith, Reinhard Lührmann

ISBN-10: 3527326065

ISBN-13: 9783527326068

ISBN-10: 3527636773

ISBN-13: 9783527636778

The booklet comprises significant components: The first one presents a quick theoretical
introduction that offers a brief assessment of other splicing and cites key papers within the field for extra in-depth details. the second one half is a set of experimental protocols which are utilized in the field of different splicing.
Contents
1 Splicing within the RNA World
2 RNPs, Small RNAs, and miRNAs
3 RNA parts inquisitive about Splicing
4 A Structural Biology point of view of Proteins fascinated about Splicing Regulation
5 The Spliceosome in Constitutive Splicing
6 using Saccharomyces cerevisiae to review the Mechanism of pre-mRNA Splicing
7 demanding situations in Plant substitute Splicing
8 substitute Splice web site Selection
9 Integration of Splicing with Nuclear and mobile Events
10 Splicing and Disease
11 From Bedside to Bench: how one can research a Splicing Mutation
12 research of universal Splicing Problems
13 Ultracentrifugation within the research and Purification of Spliceosomes Assembled In Vitro
14 Chemical Synthesis of RNA
15 RNA Interference (siRNA, shRNA)
16 Expression and Purification of Splicing Proteins
17 Detection of RNA–Protein Complexes through Electrophoretic Mobility Shift Assay
18 practical research of enormous Exonic Sequences via Iterative In Vivo Selection
19 id of Splicing cis-Elements via an Ultra-Refined Antisense Microwalk
20 Genomic SELEX to spot RNA objectives of Plant RNA-Binding Proteins
21 Quantification of different Splice Variants
22 High-Throughput research of different Splicing by means of RT-PCR
23 tracking alterations in Plant replacement Splicing Events
24 Array Analysis
25 The CLIP way to examine Protein–RNA Interactions in Intact Cells and Tissues
26 RNA–Protein Crosslinking and Immunoprecipitation (CLIP) in Schizosaccharomyces pombe
27 id of Proteins absolute to RNA
28 Single-Cell Detection of Splicing occasions with Fluorescent Splicing Reporters
29 The coaching of HeLa mobilephone Nuclear Extracts
30 In Vitro Splicing Assays
31 meeting and Isolation of Spliceosomal Complexes In Vitro
32 research of Site-Specific RNA–Protein Interactions
33 Immunoprecipitation and Pull-Down of Nuclear Proteins
34 research of Protein (-RNA) Complexes through (Quantitative) Mass Spectrometric Analysis
35 speedy Cloning of Splicing Reporter Minigenes
36 In Vivo research of Splicing Assays
37 Coupled Promoter Splicing Systems
38 sturdy phone strains with Splicing Reporters
39 Splicing issue ChIP and ChRIP: Detection ofSplicing and Splicing components at
Genes via Chromatin Immunoprecipitation
40 Yeast Genetics to enquire the functionality of center Pre-mRNA Splicing Factors
41 research of HIV-1 RNA Splicing
42 In Vivo research of Plant Intron Splicing
43 amendment State-Specific Antibodies
44 research of other Splicing in Drosophila Genetic Mosaics
45 Antisense Derivatives of U7 Small Nuclear RNA as Modulators of Pre-mRNA Splicing
46 Screening for replacement Splicing Modulators
47 Use of Oligonucleotides to alter Splicing
48 altering signs to the Spliceosome
49 review of Splicing correct Databases
50 research of RNA Transcripts by way of High-Throughput RNA Sequencing
51 identity of Splicing issue goal Genes by means of High-Throughput Sequencing
52 Bioinformatic research of Splicing Events
53 research of Pre-mRNA Secondary buildings and replacement Splicing
54 constitution Prediction for then again Spliced Proteins
55 Comparative Genomics equipment for the Prediction of Small RNA-Binding websites

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Additional info for Alternative pre-mRNA Splicing: Theory and Protocols

Sample text

1 Introduction: The Fascination of Alternative Pre-mRNA Splicing The genetic information is stored in DNA, which is transferred from one generation to the next. During the life of a cell, this DNA information is retrieved as RNA. Whereas DNA is chemically very stable and therefore well suited to archive the genetic information, RNA is chemically more reactive, and thus unstable. Therefore, with the exception of RNA viruses, RNA does not store the genetic information but rather acts as an intermediate between DNA and proteins.

J 2 RNPs, Small RNAs, and miRNAs Michaela Beitzinger and Gunter Meister Key Concepts . RNAs are almost always associated with proteins, forming RNA–protein complexes. RNA functions range from catalytic activity to pure structural scaffolding functions. Short RNAs appear to be crucial for gene regulation. RNA is processed into smaller RNA fragments that can have functions. 1 Introduction The complete sequencing of the human genome revealed that only about 2% of the genetic information is covered by protein-coding genes [1].

KGaA. Published 2012 by Wiley-VCH Verlag GmbH & Co. KGaA. 1 Overview of different cellular small RNPs. RNP Small RNA Function Reference(s) snRNP scaRNP snoRNP Telomerase miRNP/RISC miRNP/RISC RISC piRNP RITS RNase P SRP 7SK RNP tiRNP snRNA scaRNA snoRNA snoRNA, TER miRNA snoRNA-derived miRNAs siRNA piRNA siRNA (rasiRNAs) RNase P RNA 7SL RNA 7SK RNA tiRNA Splicing, major components of the spliceosome snRNA modification rRNA processing, pre-polysomes Telomere length regulation Sequence-specific gene silencing Sequence-specific gene silencing RNA interference Gene silencing, gametogenesis, transposon silencing Heterochromatin formation t-RNA maturation, transcription of tRNAs, rRNAs, snRNAs Transport of nascent proteins to the ER for further processing Regulation of transcription elongation Regulation of transcription initiation [31] [9] [9] [8] [19] [10] [13] [17,25] [18] [5–7] [26,27] [28,29] [32] In eukaryotic cells, noncoding RNA-containing RNPs are required for mRNA maturation, as well as for transcription and translation.

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Alternative pre-mRNA Splicing: Theory and Protocols by Stefan Stamm, Chris Smith, Reinhard Lührmann


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