Ribozymes and siRNA protocols
Ribozymes and siRNA protocols

Author: Mouldy Sioud

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 623

ISBN-13: 1592597467

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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

Published: 2004-03-05

Total Pages: 624

ISBN-13: 9781588292261

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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.

Antisense and Ribozyme Methodology
Antisense and Ribozyme Methodology

Author: Ian Gibson

Publisher: John Wiley & Sons

Published: 2008-09-26

Total Pages: 92

ISBN-13: 3527612521

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Antisense and ribozymes have a relatively short yet successful history as research tools in gene expression studies, and thus are considered as having high potential reagents in treating viral infections and cancer. This laboratory companion provides detailed information on the potential, advantages and limitations of this methodology. It critically discusses potential pitfalls, presents strategies for choosing targets and delivery systems, so as to allow the selection of the optimum methodology for achieving fast and reliable experimental success with any human or other biological system. For researchers, technicians and advanced graduates in experimental medicine, molecular and cell biology.

Target Discovery and Validation Reviews and Protocols
Target Discovery and Validation Reviews and Protocols

Author: Mouldy Sioud

Publisher: Springer Science & Business Media

Published: 2008-02-04

Total Pages: 360

ISBN-13: 1597451657

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Target discovery is a field that has existed for several years but is so vibrant today because of the recent progress in our understanding of the molecular mechanisms of many human diseases and the technical advances in target identification and validation. More sophisticated gene profiling technologies, such as DNA microarrays and serial analysis of gene expression, permit rapid identification of lead targets. Moreover, analysis of gene networks in living organisms allows the identification of target genes that operate in defined physiological pathways. With the sequencing of several genomes completed and the rapidly growing gene expression databases, there is now greater impetus than ever before for in silico discovery of therapeutic targets. Also, recent advances in genetic technologies have increased our ability to generate mouse models for human diseases. The implications of these genetically modified animals in drug development are several, including identification of new drug targets, predicting efficacy, and uncovering possible side effects. Together, these recent technical advances should allow researchers to make the most informed choice early and advance the chosen targets toward clinical studies. Regarding cancers, any difference between a cancer and a normal cell could potentially be exploited as a therapeutic target. The hope is that drugs targeting specific constituents or pathways in cancer cells will provide more effective therapy, either alone or in combination with other currently used anticancer drugs. In addition to drug targets, identifying new target antigens remains as much of a challenge as improving tumor vaccines already in the clinic.

Therapeutic Applications of Ribozymes
Therapeutic Applications of Ribozymes

Author: Kevin J. Scanlon

Publisher: Springer Science & Business Media

Published: 1998-06-23

Total Pages: 488

ISBN-13:

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In Therapeutic Applications of Ribozymes, expert laboratory scientists describe in detail their methodologies for constructing ribozymes designed to elucidate the role of specific genes as key routes to the development of novel therapies for a wide variety of diseases. The authors review the many sites targeted with ribozymes in various diseases and provide specific accounts of the practical techniques required for the proper use of ribozymes in these systems. Their cutting-edge protocols demonstrate how to achieve ribozyme expression in distinct cellular systems, the preparation and use of high-efficiency ribozyme DNA or RNA delivery, and the studies required to prove the efficacy of ribozyme-mediated inhibition, with its concomitant effect on phenotypic parameters. Therapeutic Applications of Ribozymes contains all the practical detail necessary to realize the high promise of ribozyme technology as a significant, widely used methodology. Focused on ribozyme targets and the novel associated technology, the book immediately becomes the leading resource for all those seeking to transform this new category of therapeutic agent into efficacious clinical practice.

Handbook of RNA Biochemistry
Handbook of RNA Biochemistry

Author: Roland K. Hartmann

Publisher: John Wiley & Sons

Published: 2015-06-22

Total Pages: 1370

ISBN-13: 3527327762

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The second edition of a highly acclaimed handbook and ready reference. Unmatched in its breadth and quality, around 100 specialists from all over the world share their up-to-date expertise and experiences, including hundreds of protocols, complete with explanations, and hitherto unpublished troubleshooting hints. They cover all modern techniques for the handling, analysis and modification of RNAs and their complexes with proteins. Throughout, they bear the practising bench scientist in mind, providing quick and reliable access to a plethora of solutions for practical questions of RNA research, ranging from simple to highly complex. This broad scope allows the treatment of specialized methods side by side with basic biochemical techniques, making the book a real treasure trove for every researcher experimenting with RNA.

MicroRNA Expression Detection Methods
MicroRNA Expression Detection Methods

Author: Zhiguo Wang

Publisher: Springer

Published: 2009-12-23

Total Pages: 382

ISBN-13: 3642049281

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MicroRNAs (miRNAs), endogenous noncoding regulatory mRNAs of - nucleotides, have rapidly emerged as the central players in gene expression regulation. Owing to their ever-increasing implications in the control of various biological and pathological processes, miRNAs have now been considered novel biomarkers of various human diseases including, cancer, viral disease, cardiov- cular disorders, metabolic disturbances, etc. Particular expression pro?les have been associated with particular pathological states. Expression pro?ling of miR- NAs have therefore become extremely important not only for fundamentalists but also for clinicians. However, the methodologies used for detecting protein-coding mRNAs cannot be directly applied to miRNAs because of their small size. Over the past years, researchers have made great efforts to developing techniques suitable for miRNA detection and quanti?cation; a wide spectrum of creative and inno- tive techniques (more than 30 different methods) have been invented and validated. It has come to the time now to summarize these methods and present them in an orderly manner for better understanding and utilization of these methods to miRNA research and applications. In particular, the development of methods for quantifying circulating miRNAs opens up a fascinating opportunity for realizing miRNA as diagnostic and prognostic biomarkers of human disease. A book on this subject may help boosting up the passion of researchers to further improve the existing techniques and develop more new methods to ?t to new application needs. These considerations prompted us and urged us to undertake the work: writing a book focusing on miRNA expression detection methods.

2-D Proteome Analysis Protocols
2-D Proteome Analysis Protocols

Author: Andrew J. Link

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 585

ISBN-13: 1592595847

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With the completion of sequencing projects and the advancement of a- lytical tools for protein identification, proteomics—the study of the expressed part of the genome—has become a major region of the burgeoning field of functional genomics. High-resolution 2-D gels can reveal virtually all p- teins present in a cell or tissue at any given time, including posttranslationally modified proteins. Changes in the expression and structure of most cellular proteins caused by differentiation or external stimuli can be displayed and eventually identified using 2-D protein gels. 2-D Proteome Analysis Protocols covers all aspects of the use of 2-D protein electrophoresis for the analysis of biological problems. The contri- tors include many of the leaders in the fields of biochemistry and analytical chemistry who were instrumental in the development of high-resolution 2-D gels, immobilized pH gradients, computer analysis, and mass spectromet- based protein identification methodologies. This book is intended as a benchtop manual and guide both for novices to 2-D gels and for those aficionados who wish to try the newer techniques. Any group using protein biochemistry—especially in the fields of molecular biology, biochemistry, microbiology, and cell biology—should find this book eminently useful. 2-D Proteome Analysis Protocols takes the researcher through the c- plete process of working with 2-D protein gels from making the protein - tract to finally identifying the proteins of interest. It includes protocols for generating 2-D protein extracts from most of the standard model organisms, including bacteria, yeast, nematode, Drosophila, plants, mouse, and human.

Polyamine Protocols
Polyamine Protocols

Author: David M. L. Morgan

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 173

ISBN-13: 1592595650

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A unique collection of hands-on enzyme assay techniques to study polyamines and their function. The techniques range from assay methods for enzymes of polyamine biosynthesis and catabolism to measurements of polyamines, polyamine transport, and polyamine effects on cell growth. The methods are presented by leading researchers who have perfected them to a high art, and include clear, step-by-step instructions with numerous hints and tips to ensure readily reproducible results.

Immunochemical Protocols
Immunochemical Protocols

Author: John Pound

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 501

ISBN-13: 1592592570

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This much anticipated second edition provides a user-friendly, up-to-date handbook of reliable immunochemical techniques optimized for molecular biologists. It covers the breadth of relevant established methods from protein blotting and immunoassays through to visualization of cellular antigens and in situ hybridization, each with their latest refinements. Protocols for the production and purification of important classes of immunochemical reagents are also provided, including "conventional" and recombinant antibodies, fusion proteins and their various conjugates. This book will open the door to a new generation of immunochemical reagents with exciting possibilities.