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RNA Silencing

Methods and Protocols

  • Book
  • © 2005

Overview

Editors:
  1. Gordon G. Carmichael

    1. Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington

    View editor publications

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Part of the book series: Methods in Molecular Biology (MIMB, volume 309)

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Table of contents (19 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. Identification of Components of RNAi Pathways Using the Tandem Affinity Purification Method

    • Mikiko C. Siomi, Haruhiko Siomi
    Pages 1-9

  3. Separation of Drosophila RNA Silencing Complexes by Native Gel Electrophoresis

    • John W. Pham, Erik J. Sontheimer
    Pages 11-16

  4. Preparation and Analysis of Drosha

    • Yoontae Lee, V. Narry Kim
    Pages 17-28

  5. RNA Interference in Caenorhabditis elegans

    • Nathaniel R. Dudley, Bob Goldstein
    Pages 29-38

  6. Down-Regulating Gene Expression by RNA Interference in Trypanosoma brucei

    • Christine E. Clayton, Antonio M. Estévez, Claudia Hartmann, Vincent P. Alibu, Mark Field, David Horn
    Pages 39-59

  7. Analysis of Double-Stranded RNA and Small RNAs Involved in RNA-Mediated Transcriptional Gene Silencing

    • M. Florian Mette, Werner Aufsatz, Tatsuo Kanno, Lucia Daxinger, Philipp Rovina, Marjori Matzke et al.
    Pages 61-82

  8. Strategies for the Design of Random siRNA Libraries and the Selection of anti-GFP siRNAs

    • Mouldy Sioud, Sébastien Wälchli
    Pages 83-91

  9. Silencing Gene Expression with Dicer-Generated siRNA Pools

    • Jason W. Myers, James E. Ferrell Jr.
    Pages 93-196

  10. Inhibition of Gene Expression In Vivo Using Multiplex siRNA

    • Sung-Suk Chae, Timothy Hla
    Pages 197-203

  11. Gene Silencing by a DNA Vector-Based RNAi Technology

    • Guangchao Sui, Yang Shi
    Pages 205-218

  12. Plasmid-Based RNA Interference

    • Scott Q. Harper, Beverly L. Davidson
    Pages 219-235

  13. siRNA Delivery In Vivo

    • Mouldy Sioud
    Pages 237-249

  14. Peptide-Based Strategy for siRNA Delivery into Mammalian Cells

    • Federica Simeoni, May C. Morris, Frederic Heitz, Gilles Divita
    Pages 251-260

  15. Lentiviral Vector Delivery of siRNA and shRNA Encoding Genes into Cultured and Primary Hematopoietic Cells

    • Mingjie Li, John J. Rossi
    Pages 261-272

  16. Exon-Specific RNA Interference

    • Alicia M. Celotto, Joo-Won Lee, Brenton R. Graveley
    Pages 273-282

  17. Immunoprecipitation of MicroRNPs and Directional Cloning of MicroRNAs

    • Elisavet Maniataki, Maria Dels Angels De Planell Saguer, Zissimos Mourelatos
    Pages 283-294

  18. Detection of MicroRNAs and Assays to Monitor MicroRNA Activities In Vivo and In Vitro

    • Marianthi Kiriakidou, Peter Nelson, Stella Lamprinaki, Anup Sharma, Zissimos Mourelatos
    Pages 295-310

  19. High-Throughput Analysis of MicroRNA Gene Expression Using Sensitive Probes

    • Mouldy Sioud, Øystein Røsok
    Pages 311-320

  20. Modification of Human U1 snRNA for Inhibition of Gene Expression at the Level of Pre-mRNA

    • Peng Liu, Mary Louise Stover, Alexander Lichtler, David W. Rowe
    Pages 321-331
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Keywords

About this book

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 protocols 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.

Reviews

From the reviews:

"RNA Silencing: Methods and Protocols 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)

Editors and Affiliations

  • Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington

    Gordon G. Carmichael

Bibliographic Information

  • Book Title: RNA Silencing

  • Book Subtitle: Methods and Protocols

  • Editors: Gordon G. Carmichael

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1385/1592599354

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 2005

  • Hardcover ISBN: 978-1-58829-436-4Published: 01 June 2005

  • Softcover ISBN: 978-1-61737-568-2Published: 19 November 2010

  • eBook ISBN: 978-1-59259-935-6Published: 04 February 2008

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: X, 338

  • Number of Illustrations: 74 b/w illustrations

  • Topics: Cell Biology

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