Ribozyme Protocols

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Ribozymes and siRNA protocols

Author : Mouldy Sioud
Publisher : Springer Science & Business Media
ISBN 13 : 1592597467
Total Pages : 623 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Ribozymes and siRNA protocols by : Mouldy Sioud

Download or read book Ribozymes and siRNA protocols written by Mouldy Sioud and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 623 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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

Author : Mouldy Sioud
Publisher : Humana Press
ISBN 13 : 9781588292261
Total Pages : 624 pages
Book Rating : 4.2/5 (922 download)

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Book Synopsis Ribozymes and siRNA protocols by : Mouldy Sioud

Download or read book Ribozymes and siRNA protocols written by Mouldy Sioud and published by Humana Press. This book was released on 2004-03-05 with total page 624 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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

Author : Ian Gibson
Publisher : John Wiley & Sons
ISBN 13 : 3527612521
Total Pages : 92 pages
Book Rating : 4.5/5 (276 download)

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Book Synopsis Antisense and Ribozyme Methodology by : Ian Gibson

Download or read book Antisense and Ribozyme Methodology written by Ian Gibson and published by John Wiley & Sons. This book was released on 2008-09-26 with total page 92 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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

Author : Mouldy Sioud
Publisher : Springer Science & Business Media
ISBN 13 : 1597451657
Total Pages : 360 pages
Book Rating : 4.5/5 (974 download)

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Book Synopsis Target Discovery and Validation Reviews and Protocols by : Mouldy Sioud

Download or read book Target Discovery and Validation Reviews and Protocols written by Mouldy Sioud and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 360 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Handbook of RNA Biochemistry

Author : Roland K. Hartmann
Publisher : John Wiley & Sons
ISBN 13 : 3527327762
Total Pages : 1370 pages
Book Rating : 4.5/5 (273 download)

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Book Synopsis Handbook of RNA Biochemistry by : Roland K. Hartmann

Download or read book Handbook of RNA Biochemistry written by Roland K. Hartmann and published by John Wiley & Sons. This book was released on 2015-06-22 with total page 1370 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Therapeutic Applications of Ribozymes

Author : Kevin J. Scanlon
Publisher : Springer Science & Business Media
ISBN 13 :
Total Pages : 488 pages
Book Rating : 4.3/5 (91 download)

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Book Synopsis Therapeutic Applications of Ribozymes by : Kevin J. Scanlon

Download or read book Therapeutic Applications of Ribozymes written by Kevin J. Scanlon and published by Springer Science & Business Media. This book was released on 1998-06-23 with total page 488 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

MicroRNA Expression Detection Methods

Author : Zhiguo Wang
Publisher : Springer
ISBN 13 : 3642049281
Total Pages : 382 pages
Book Rating : 4.6/5 (42 download)

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Book Synopsis MicroRNA Expression Detection Methods by : Zhiguo Wang

Download or read book MicroRNA Expression Detection Methods written by Zhiguo Wang and published by Springer. This book was released on 2009-12-23 with total page 382 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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

Author : Andrew J. Link
Publisher : Springer Science & Business Media
ISBN 13 : 1592595847
Total Pages : 585 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis 2-D Proteome Analysis Protocols by : Andrew J. Link

Download or read book 2-D Proteome Analysis Protocols written by Andrew J. Link and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 585 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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

Author : David M. L. Morgan
Publisher : Springer Science & Business Media
ISBN 13 : 1592595650
Total Pages : 173 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Polyamine Protocols by : David M. L. Morgan

Download or read book Polyamine Protocols written by David M. L. Morgan and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 173 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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

Author : John Pound
Publisher : Springer Science & Business Media
ISBN 13 : 1592592570
Total Pages : 501 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Immunochemical Protocols by : John Pound

Download or read book Immunochemical Protocols written by John Pound and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 501 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Glycoanalysis Protocols

Author : Elizabeth F. Hounsell
Publisher : Springer Science & Business Media
ISBN 13 : 1592595626
Total Pages : 263 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Glycoanalysis Protocols by : Elizabeth F. Hounsell

Download or read book Glycoanalysis Protocols written by Elizabeth F. Hounsell and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 263 pages. Available in PDF, EPUB and Kindle. Book excerpt: Now fully updated and considerably expanded, Glycoanalysis Protocols, 2nd ed., makes available to all protein scientists, and particularly those working with today's pharmaceuticals, the most advanced and reproducible glycoanalysis techniques currently in use. Developed by highly experienced carbohydrate chemists, biochemists, and physical chemists, these detailed, up-to-date, and proven analytical techniques cover the areas of glycoprotein macromolecular structural analysis, oligosaccharide profiling, lipid conjugate characterization, microorganism structure determination, and proteoglycan function. Special attention has been given to advanced analytical techniques in biotechnology during the production of recombinant glycoproteins and other therapeutics. Hailed as "indispensable" in its first edition, Glycoanalysis Protocols, 2nd ed., continues with vital, time-tested techniques addressing the needs of both biomedical researchers and protein macromolecular structural chemists. It will well serve all those starting work on the analysis of glycoproteins, as well as more experienced investigators seeking to augment their expertise.

Cytochrome P450 Protocols

Author : Ian R. Phillips
Publisher : Springer Science & Business Media
ISBN 13 : 1592595804
Total Pages : 473 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Cytochrome P450 Protocols by : Ian R. Phillips

Download or read book Cytochrome P450 Protocols written by Ian R. Phillips and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 473 pages. Available in PDF, EPUB and Kindle. Book excerpt: In Cytochrome P450 Protocols, Ian Phillips and Elizabeth Shephard assemble a comprehensive collection of cutting-edge techniques for the investigation of cytochromes P450. Described in detail by hands-on experimentalists for easy reproducibility, these methods include spectral analysis, purification and enzymatic assays, expression in heterologous systems, and the production and use of antibodies, as well as methods for quantification of gene expression, transfection of hepatocytes, and for the investigation of DNA-protein interactions and genetic polymorphisms. In addition, because of the growing importance of in vitro systems in pharmacological toxicology, the book contains techniques for the culture of rodent and human hepatocytes and human epidermis. Cytochrome P450 induction as a biomarker for environmental pollution and the generation of mice with targeted gene disruptions complete this exhaustive collection of core techniques. Cytochrome P450 Protocols includes in one volume both state-of-the-art and classic methods that have not been superseded but remain extremely useful. The collection provides both novice and experienced researchers across many fields-toxicology, pharmacology, environmental biology, biochemistry, and molecular biology-all the tools needed to elucidate the crucial biological role played by cytochromes P450 in the metabolism of therapeutic drugs, chemical carcinogens, and environmental pollutants.

Transmembrane Signaling Protocols

Author : Dafna Bar-Sagi
Publisher : Springer Science & Business Media
ISBN 13 : 0896034887
Total Pages : 631 pages
Book Rating : 4.8/5 (96 download)

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Book Synopsis Transmembrane Signaling Protocols by : Dafna Bar-Sagi

Download or read book Transmembrane Signaling Protocols written by Dafna Bar-Sagi and published by Springer Science & Business Media. This book was released on 1998 with total page 631 pages. Available in PDF, EPUB and Kindle. Book excerpt: This collection of practical, cutting-edge techniques for the study of cell signaling provides detailed, step-by-step instructions, helpful notes, and troubleshooting tips that make even the most powerful of the newest techniques readily reproducible. The protocols presented include the use of peptide libraries to study transmembrane signaling; the use of single-cell assays to analyze signal transduction pathways; the reconstitution of signaling complexes; methods for analyzing protein-protein interactions, and more. Introductory reviews explain the basic theory and enable researchers new to the area to rapidly gain understanding, as well as command of the practical knowledge and expertise afforded by the protocols. Transmembrane Signaling Protocols makes available to all researchers the many state-of-the-art techniques that have recently led to landmark discoveries in transmembrane signaling.

Protein Targeting Protocols

Author : Roger A. Clegg
Publisher : Springer Science & Business Media
ISBN 13 : 1592595723
Total Pages : 331 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Protein Targeting Protocols by : Roger A. Clegg

Download or read book Protein Targeting Protocols written by Roger A. Clegg and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 331 pages. Available in PDF, EPUB and Kindle. Book excerpt: It is by no means a revelation that proteins are not uniformly distributed throughout the cell. As a result, the idea that protein molecules, because of the specificity with which they can engage in interactions with other proteins, may be aimed—via these interactions—at a restricted target, is a fundamental one in contemporary molecular life sciences. The target may be variously c- ceived as a specific molecule, a group of molecules, a structure, or a more generic type of intracellular environment. Because the concept of protein targeting is intuitive rather than expl- itly defined, it has been variously used by different groups of researchers in cell biology, biochemistry, and molecular biology. For those working in the field of intracellular signaling, an influential introduction to the topic was the seminal article by Hubbard & Cohen (TIBS [1993] 18, 172–177), which was based on the work of Cohen’s laboratory on protein phosphatases. Sub- quently, the ideas that they discussed have been further developed and extended by many workers to other key intermediaries in intracellular sign- ing, including protein kinases and a great variety of modulator and adaptor proteins.

Plant Virology Protocols

Author : Gary D. Foster
Publisher : Springer Science & Business Media
ISBN 13 : 1592595669
Total Pages : 557 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Plant Virology Protocols by : Gary D. Foster

Download or read book Plant Virology Protocols written by Gary D. Foster and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 557 pages. Available in PDF, EPUB and Kindle. Book excerpt: The aim of Plant Virology Protocols is to provide a source of infor- tion to guide the reader through the wide range of methods involved in gen- ating transgenic plants that are resistant to plant viruses. To this end, we have commissioned a wide-ranging list of chapters that will cover the methods required for: plant virus isolation; RNA extraction; cloning coat p- tein genes; introduction of the coat protein gene into the plant genome; and testing transgenic plants for resistance. The book then moves on to treatments of the mechanisms of resistance, the problems encountered with field testing, and key ethical issues surrounding transgenic technology. Although Plant Virology Protocols deals with the cloning and expression of the coat protein gene, the techniques described can be equally applied to other viral genes and nucleotide sequences, many of which have also been shown to afford protection when introduced into plants. The coat protein has, however, been the most widely applied, and as such has been selected to illustrate the techniques involved. Plant Virology Protocols has been divided into six major sections, c- taining 55 chapters in total.

Combinatorial Peptide Library Protocols

Author : Shmuel Cabilly
Publisher : Springer Science & Business Media
ISBN 13 : 1592595715
Total Pages : 320 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Combinatorial Peptide Library Protocols by : Shmuel Cabilly

Download or read book Combinatorial Peptide Library Protocols written by Shmuel Cabilly and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 320 pages. Available in PDF, EPUB and Kindle. Book excerpt: During the course of evolution, an imbalance was created between the rate of vertebrate genetic adaptation and that of the lower forms of living organisms, such as bacteria and viruses. This imbalance has given the latter the advantage of generating, relatively quickly, molecules with unexpected structures and features that carry a threat to vertebrates. To compensate for their weakness, vertebrates have accelerated their own evolutionary processes, not at the level of whole organism, but in specialized cells containing the genes that code for antibody molecules or for T-cell receptors. That is, when an immediate requirement for molecules capable of specific interactions arose, nature has preferred to speed up the mode of Darwinian evolution in pref- ence to any other approach (such as the use of X-ray diffraction studies and computergraphic analysis). Recently, Darwinian rules have been adapted for test tube research, and the concept of selecting molecules having particular characteristics from r- dom pools has been realized in the form of various chemical and biological combinatorial libraries. While working with these libraries, we noticed the interesting fact that when combinatorial libraries of oligopeptides were allowed to interact with different selector proteins, only the actual binding sites of these proteins showed binding properties, whereas the rest of the p- tein surface seemed "inert. " This seemingly common feature of protein- having no extra potential binding sites--was probably selected during evolution in order to minimize nonspecific interactions with the surrounding milieu.

Lipase and Phospholipase Protocols

Author : Mark Doolittle
Publisher : Springer Science & Business Media
ISBN 13 : 1592595812
Total Pages : 362 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Lipase and Phospholipase Protocols by : Mark Doolittle

Download or read book Lipase and Phospholipase Protocols written by Mark Doolittle and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 362 pages. Available in PDF, EPUB and Kindle. Book excerpt: The lipases and phospholipases represent a diverse group of enzymes that are expressed in animals, plants, fungi, and bacteria. Their ubiquitous distribution among all species is a testament to the essential roles played by these enzymes in lipid storage, mobilization and metabolism, membrane homeostasis and remodeling, endocrine and immune functions, and signal tra- duction. In humans, lipases and phospholipases are also thought to contribute to complex diseases, such as atherosclerosis, obesity, arthritis, and cancer, as well as to single gene defects, such as Wolman's disease and Type I hyperlipoproteinemia. Enzymatically, the lipases are unique, since they hydrolyze substrates that are either insoluble, or only partly soluble, in aq- ous solvents; thus, enzyme catalysis takes place at a lipid-water interface. The interface comprises at least two, and often more, discrete bulk and s- face phases, in which the enzyme, substrate, and products oflipolysis disperse among these phases based on their physical properties. Furthermore, the d- tribution of these components changes continuously as lipolysis proceeds. Thus, the lipases and phospholipases are fundamentally different from any other enzyme because of the physical complexity of the environment in which catalysis occurs.