RNA silencing : methods and protocols / edited by Gordon Carmichael.

Contributor(s): Carmichael, Gordon
Material type: TextTextSeries: Methods in molecular biology (Clifton, N.J.): v. 309.Publisher: Totowa, N.J. : Humana Press, ©2005Description: 1 online resource (xi, 338 pages) : illustrationsContent type: text Media type: computer Carrier type: online resourceISBN: 1588294366; 9781588294364; 9781592599356; 1592599354Subject(s): Small interfering RNA -- Laboratory manuals | Gene silencing -- Laboratory manuals | Gene Expression Regulation -- genetics | MicroRNAs -- physiology | RNA, Small Interfering -- physiology | RNA Interference | MicroRNAs | Physiology | RNA, Small Interfering | Genetics | Gene Expression Regulation | Genetic Phenomena | Biological Science Disciplines | Gene Silencing | Biology | RNA, Small Untranslated | RNA, Antisense | RNA | RNA, Untranslated | Antisense Elements (Genetics) | Epigenesis, Genetic | Natural Science Disciplines | Phenomena and Processes | Nucleic Acids, Nucleotides, and Nucleosides | Nucleic Acids | Disciplines and Occupations | Chemicals and Drugs | SCIENCE -- Life Sciences -- Genetics & Genomics | Gene silencing | Small interfering RNA | RnaGenre/Form: Fulltext. | Index not Present. | Internet Resource. | Laboratory Manual. | Laboratory manuals. Additional physical formats: Print version:: RNA silencing.DDC classification: 572.8/65 LOC classification: QP623.5.S63 | R634 2005Online resources: Click here to access online
Contents:
Identification of components of RNAi pathways using the tandem affinity purification method / Mikiko C. Siomi and Haruhiko Siomi -- Separation of drosophila RNA silencing complexes by native gel electrophoresis / John W. Pham and Erik J. Sontheimer -- Preparation and analysis of drosha / Yoontae Lee and V. Narry Kim -- RNA interference in Caenorhabditis elegans / Nathaniel R. Dudley and Bob Goldstein -- Down-regulating gene expression by RNA interference in Trypanosoma brucei / Christine E. Clayton [and others] -- Analysis of double-stranded RNA and small RNAs involved in RNA-mediated transcriptional gene silencing / M. Florian Mette [and others] -- Strategies for the design of random siRNA libraries and the selection of anti-GFP siRNAs / Mouldy Sioud and Sebastiona Wulchli -- Silencing gene expression with dicer-generated siRNA pools / Jason W. Myers and James E. Ferrell, Jr.
Inhibition of gene expression in vivo using multiplex siRNA / Sung-Suk Chae and Timothy Hla -- Gene silencing by a DNA vector-based RNAi technology / Guangchao Sui and Yang Shi -- Plasmid-based RNA interference: construction of small-hairpin RNA expression vectors / Scott Q. Harper and Beverly L. Davidson -- SiRNA delivery in vivo / Mouldy Sioud -- Peptide-based strategy for siRNA delivery into mammalian cells / Federica Simeoni [and others] -- Lentiviral vector delivery of siRNA and shRNA encoding genes into cultured and primary hematopoietic cells / Mingjie Li and Jo Rossi -- Exon-specific RNA interference: a tool to determine the functional relevance of proteins encoded by alternatively spliced mRNAs / Alicia M. Celotto, Joo-Won Lee, and Brenton R. Graveley -- Immunoprecipitation of microRNPs and directional cloning of microRNAs / Elsa Maniataki, Maria Dels Angels de Planell Saguer, and Zissimos Mourelatos.
Detection of microRNAs and assays to monitor microRNA activities in vivo and in vitro / Marianthi Kiriakidou [and others] -- High throughput analysis of microRNA gene expression using sensitive cDNA probes / Mouldy Sioud and Øystein Røsok -- Modification of human U1 snRNA for inhibition of gene expression at the level of pre-mRNA / Peng Liu [and others].
Summary: The past decade has witnessed a true revolution in our understanding of how RNA molecules can reduce or silence gene expression, a process that holds great promise in basic research and for antiviral and anticancer applications. In RNA Silencing: Methods and Protocols, many of the most notable and distinguished researchers in the field describe in step-by-step detail their best methods for the design, preparation, and use of RNAs to reduce 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. Comprehensive and state-of-the-art, RNA Silencing: Methods and Protocols offers today's laboratory investigators a host of optimized and validated techniques to study RNA silencing and translate its concepts into successful practical applications.
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Includes bibliographical references and index.

Identification of components of RNAi pathways using the tandem affinity purification method / Mikiko C. Siomi and Haruhiko Siomi -- Separation of drosophila RNA silencing complexes by native gel electrophoresis / John W. Pham and Erik J. Sontheimer -- Preparation and analysis of drosha / Yoontae Lee and V. Narry Kim -- RNA interference in Caenorhabditis elegans / Nathaniel R. Dudley and Bob Goldstein -- Down-regulating gene expression by RNA interference in Trypanosoma brucei / Christine E. Clayton [and others] -- Analysis of double-stranded RNA and small RNAs involved in RNA-mediated transcriptional gene silencing / M. Florian Mette [and others] -- Strategies for the design of random siRNA libraries and the selection of anti-GFP siRNAs / Mouldy Sioud and Sebastiona Wulchli -- Silencing gene expression with dicer-generated siRNA pools / Jason W. Myers and James E. Ferrell, Jr.

Inhibition of gene expression in vivo using multiplex siRNA / Sung-Suk Chae and Timothy Hla -- Gene silencing by a DNA vector-based RNAi technology / Guangchao Sui and Yang Shi -- Plasmid-based RNA interference: construction of small-hairpin RNA expression vectors / Scott Q. Harper and Beverly L. Davidson -- SiRNA delivery in vivo / Mouldy Sioud -- Peptide-based strategy for siRNA delivery into mammalian cells / Federica Simeoni [and others] -- Lentiviral vector delivery of siRNA and shRNA encoding genes into cultured and primary hematopoietic cells / Mingjie Li and Jo Rossi -- Exon-specific RNA interference: a tool to determine the functional relevance of proteins encoded by alternatively spliced mRNAs / Alicia M. Celotto, Joo-Won Lee, and Brenton R. Graveley -- Immunoprecipitation of microRNPs and directional cloning of microRNAs / Elsa Maniataki, Maria Dels Angels de Planell Saguer, and Zissimos Mourelatos.

Detection of microRNAs and assays to monitor microRNA activities in vivo and in vitro / Marianthi Kiriakidou [and others] -- High throughput analysis of microRNA gene expression using sensitive cDNA probes / Mouldy Sioud and Øystein Røsok -- Modification of human U1 snRNA for inhibition of gene expression at the level of pre-mRNA / Peng Liu [and others].

Print version record.

The past decade has witnessed a true revolution in our understanding of how RNA molecules can reduce or silence gene expression, a process that holds great promise in basic research and for antiviral and anticancer applications. In RNA Silencing: Methods and Protocols, many of the most notable and distinguished researchers in the field describe in step-by-step detail their best methods for the design, preparation, and use of RNAs to reduce 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. Comprehensive and state-of-the-art, RNA Silencing: Methods and Protocols offers today's laboratory investigators a host of optimized and validated techniques to study RNA silencing and translate its concepts into successful practical applications.

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