Ribozymes and siRNA protocols

Ribozymes and siRNA protocols
Author: Mouldy Sioud
Publisher: Springer Science & Business Media
Total Pages: 623
Release: 2008-02-03
Genre: Science
ISBN: 1592597467

In this completely updated and expanded edition of a classic bench manual, hands-on experts take advantage of the latest advances in ribozyme, DNAzyme, hammerhead ribozymes and derivatives, and RNA interference technologies to describe in detail the exciting and successful methods now available for gene inactivation in vitro and in vivo. Their optimized techniques employ hairpin ribozymes, DNAzymes, hammerhead ribozymes and derivatives, group I intron ribozymes, RNase P ribozymes, and siRNAs, as well as general methods for RNA structure analysis, delivery of oligonucleotides, and gene therapy. Also provided are novel methods for identifying accessible cellular mRNA sites; group I intron and RNAse P ribozyme protocols for effective design, selection, and therapeutic applications; and the latest RNAi methods for sequence-specific gene silencing in a wide variety of organisms. Additional techniques cover the analysis of ribozyme structures and conformational transitions using nucleotide analog interference mapping and fluorescence resonance energy transfer, the use of ribozymes in clinical and gene therapy, and the use of ribozymes and DNAzymes in rodent models of human disease. Each proven protocol includes a background introduction outlining the principle behind the technique, step-by-step instructions, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and up-to-date, Ribozymes and siRNA Protocols details for experienced and novice investigators alike the many exciting advances in our understanding of nucleic acid enzymes, as well as demonstrating how they may be used to analyze gene function and target validation, and to productively develop novel therapeutics for human diseases.

Ribozymes and siRNA protocols

Ribozymes and siRNA protocols
Author: Mouldy Sioud
Publisher: Humana Press
Total Pages: 624
Release: 2004-03-05
Genre: Science
ISBN: 9781588292261

In this completely updated and expanded edition of a classic bench manual, hands-on experts take advantage of the latest advances in ribozyme, DNAzyme, and RNA interference technologies to describe in detail the exciting and successful methods now available for gene inactivation in vitro and in vivo. Their optimized techniques employ hairpain ribozymes, DNAzymes, hammerhead ribozymes and derivatives, group I intron ribozymes, Rnase P ribozymes, and siRNAs, as well as general methods for RNA structure analysis, delivery of oligonucleotides, and gene therapy. Also provided are novel methods for identifying accessible cellular mRNA sites; group I intron and RNAse P ribozymes protocols for effective design, selection, and therapeutic applications; and the latest RNAi methods for sequencing-specific gene silencing in a wide variety of organisms. Comprehensive and up-to-date, Ribozymes and siRNA Protocols synthesizes for experienced and novice investigators alike the exciting advances in understanding nucleic acid enzymes and demonstrates how they may be used to analyze gene function and target validation, and to productively develop new therapeutics for human diseases.

Ribozyme Protocols

Ribozyme Protocols
Author: Philip C. Turner
Publisher: Humana
Total Pages: 492
Release: 2013-09-20
Genre: Science
ISBN: 9781489942852

The purpose of Ribozyme Protocols is to provide a helpful compilation of protocols that will be of use—^not only to those with some experience of ribozymes—^but also to those wishing to use ribozymes for the first time. Although it is usually impossible to cover every aspect of a scientific field, I believe this book approaches that ideal and should help all readers perform meaningful experiments using ribozymes. To design ribozymes, one must consider whether the target site will be accessible; this task can be facilitated by using computer programs that pre dict the folding of the target RNA. Such programs are detailed in Chapters 2 and 3. If the chosen target is an RNA virus that can mutate rapidly, it makes sense to consider those parts of the genome that are least likely to change during viral replication. An example of how this can be done is described in Chapter 4. Although computer analysis may be a useful starting point to select tar get sites, there seems, at the moment, to be no guarantee that any particular chosen site will be efficiently cleaved. Some workers have deliberately bypassed this problem by using libraries of ribozyme sequences and by select ing those that actually hybridize to and/or cleave the target; these methods are described in Chapters 5 and 6.

RNA Therapeutics

RNA Therapeutics
Author: Mouldy Sioud
Publisher: Humana
Total Pages: 526
Release: 2016-08-23
Genre: Science
ISBN: 9781493961511

Central to the synthesis of proteins, the performance of catalysis, and many other physiological processes, the aberrant expression of which can be linked to human diseases including cancers, RNA has proven to be key target for therapeutics as well as a tool for therapy. In RNA Therapeutics: Function, Design, and Delivery, expert contributors from a broad spectrum of scientific backgrounds highlight the roles that messenger RNAs and small RNAs can play in biology and medicine. While covering the five major RNA-based drugs, namely the use of ribozymes to cleave and/or correct mRNA transcript, the use of siRNA for targeted silencing of gene transcripts, the use of aptamers, like short RNA molecules, for neutralizing the protein functions, the use mRNA-transfected DCs to activate immune system against tumor cells, as well as the use of RNA to reprogram T and/or DC cell function, this extensive volume brings together the fields of coding (mRNA) and non-coding RNA such as ribozymes, RNAse P, siRNAs, and miRNAs into one convenient source. Written in the highly successful Methods in Molecular BiologyTM series format, the cutting-edge protocol chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and practical tips on troubleshooting and avoiding known pitfalls. Also, the book contains several excellent reviews for teaching purposes. Authoritative and comprehensive, RNA Therapeutics: Function, Design, and Delivery provides key models and tools which will assist researchers in increasing our understanding of RNA functions, modifications, and their involvement in diseases in order to lead to the design of vital new RNA-based therapeutics.

Ribozymes

Ribozymes
Author: Jörg S. Hartig
Publisher: Humana Press
Total Pages: 480
Release: 2012-02-09
Genre: Science
ISBN: 9781617795442

Over the past decade, there have been a variety of fundamental insights into ribozymes which have changed the perspective on RNA catalysis. In, Ribozymes: Methods and Protocols, researchers in the field detail many of the protocols and techniques which are now commonly used to study individual ribozymes and the characterization of ribozymes. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Ribozymes: Methods and Protocols seeks to aid scientists in further understanding the catalytically active RNAs .

Therapeutic Applications of Ribozymes and Riboswitches

Therapeutic Applications of Ribozymes and Riboswitches
Author: Daniel Lafontaine
Publisher:
Total Pages: 278
Release: 2014
Genre: Catalytic RNA
ISBN: 9781627037303

In Therapeutic Applications of Ribozymes and Riboswitches: Methods and Protocols, expert researchers in the field provide a complete overview of protocols used in the development of RNA molecule as drugs and drug target. These include methods and protocols on recent and precise RNA molecule approaches or studies in the development of an RNA therapeutic tool. We are convinced that these methods will help researchers from various domains of life sciences, including clinicians, biochemists and virologists. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Therapeutic Applications of Ribozymes and Riboswitches: Methods and Protocols seeks to aid scientists in the further development of new therapeutic applications implicating RNA molecules or directly targeting harmful RNA.

Target Discovery and Validation Reviews and Protocols

Target Discovery and Validation Reviews and Protocols
Author: Mouldy Sioud
Publisher: Springer Science & Business Media
Total Pages: 359
Release: 2008-02-05
Genre: Medical
ISBN: 1597452084

These volumes review the most current methods for drug target discovery and validation. They explore how recent improvement in understanding the molecular mechanisms of human pathology is impacting drug target discovery in the laboratory and in real therapeutics, specifically for cancers and autoimmune disorders. This book provides a thorough review of the most cutting-edge methods available for each step in drug target identification, validation, and clinical application.

RNA Towards Medicine

RNA Towards Medicine
Author: Volker A. Erdmann
Publisher: Springer Science & Business Media
Total Pages: 463
Release: 2006-02-04
Genre: Science
ISBN: 3540272623

Developments over the past few years have revealed the remarkable versatility of RNA in any compartment of the cell, tasks that had been thought to be exclusively in the realm of proteins and even beyond. The chapters in this book written by leading investigators in the field provide insight into various promising avenues where RNA and nucleic acid derivatives including antisense RNAs, such as siRNA, miRNAs, amplification/selection (SELEX) generated aptamers as well as ribozymes are at the threshold of impacting medicine.

Cytokine Protocols

Cytokine Protocols
Author: Marc De Ley
Publisher: Springer Science & Business Media
Total Pages: 248
Release: 2008-02-03
Genre: Medical
ISBN: 1592596673

A collection of biochemical, cellular, and molecular techniques for unraveling and quantifying the events occurring between the initial contact of a cytokine at the membrane receptor and the eventual activation of gene transcription. The techniques used include the generation of transfectants, the immunohistochemical detection of cytokines in tissue sections, and optimized staining for cytoplasmic detection. Highlights include RT-PCR of small amounts of mRNA, in situ hybridization, biosensor analysis, measurement of biological activities and standardization, immunohistochemical and single-cell detection, and receptor isolation, characterization, and crystallization. Enjoy a quick and smooth introduction to the key methods used in cytokine research Use readily reproducible techniques that ensure successful experimental results Employ antisense-RNA, RT-PCR of small amounts of mRNA, and in situ hybridization.

Drosophila Cytogenetics Protocols

Drosophila Cytogenetics Protocols
Author: Daryl S. Henderson
Publisher: Springer Science & Business Media
Total Pages: 467
Release: 2008-02-03
Genre: Science
ISBN: 1592596657

Leading drosophilists describe in step-by-step detail all the essential techniques for studying Drosophila chromosomes and suggest new avenues for scientific exploration. The chapters emphasize specimen preparation (from dissection to mounting) and cover both polytene and mitotic/meiotic chromosomes in depth. Each fully tested and readily reproducible protocol offers a background introduction, equipment and reagent lists, and tips on troubleshooting and avoiding pitfalls. A cutting-edge FISH and immunolocalization technique will be important for discovering how DNA sequence influences higher-order chromosome architecture and ultimately gene expression.