RNA Silencing: Methods and Protocols / Edition 1

by Gordon Carmichael

Overview

A collection of readily reproducible methods for the design, preparation, and use of RNAs for silencing gene expression in cells and organisms. The techniques range widely and include methods addressing the biochemical aspects of the silencing machinery, RNA silencing in non-mammalian organisms, and the in vivo delivery of siRNAs and silencing vectors. There are also techniques for designing, preparing, and using RNAs to silence gene expression, for fine-tuning regulation by targeting specific isoforms of a given gene, and for the study and use of microRNAs. The prools follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Product Details

ISBN-13:	9781617375682
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/19/2010
Series:	Methods in Molecular Biology , #309
Edition description:	Softcover reprint of hardcover 1st ed. 2005
Pages:	338
Product dimensions:	5.98(w) x 9.02(h) x 0.03(d)

Table of Contents

Identification of Components of RNAi Pathways Using the Tandem Affinity Purification Method.- Separation of Drosophila RNA Silencing Complexes by Native Gel Electrophoresis.- Preparation and Analysis of Drosha.- RNA Interference in Caenorhabditis elegans.- Down-Regulating Gene Expression by RNA Interference in Trypanosoma brucei.- Analysis of Double-Stranded RNA and Small RNAs Involved in RNA-Mediated Transcriptional Gene Silencing.- Strategies for the Design of Random siRNA Libraries and the Selection of anti-GFP siRNAs.- Silencing Gene Expression with Dicer-Generated siRNA Pools.- Inhibition of Gene Expression In Vivo Using Multiplex siRNA.- Gene Silencing by a DNA Vector-Based RNAi Technology.- Plasmid-Based RNA Interference.- siRNA Delivery In Vivo.- Peptide-Based Strategy for siRNA Delivery into Mammalian Cells.- Lentiviral Vector Delivery of siRNA and shRNA Encoding Genes into Cultured and Primary Hematopoietic Cells.- Exon-Specific RNA Interference.- Immunoprecipitation of MicroRNPs and Directional Cloning of MicroRNAs.- Detection of MicroRNAs and Assays to Monitor MicroRNA Activities In Vivo and In Vitro.- High-Throughput Analysis of MicroRNA Gene Expression Using Sensitive Probes.- Modification of Human U1 snRNA for Inhibition of Gene Expression at the Level of Pre-mRNA.

Editorial Reviews

From the reviews:

"RNA Silencing: Methods and Prools is a comprehensive and user-friendly reference and worth recommending. Researchers who are interested in using siRNA as a major method to do gene knockdown experiments in vitro to study gene function instead of using time-consuming gene targeting in mice and those who are interested in studying the functions of genes in the development of cell defense against viruses will find this book an excellent resource." (Weimin He, In Vitro Cellular & Developmental Biology - Animal, Vol. 42, May and June, 2006)

From the Publisher