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Author: Philip C. Turner Publisher: Humana ISBN: 9781489942852 Category : Science Languages : en Pages : 492
Book Description
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.
Author: Philip C. Turner Publisher: Humana ISBN: 9781489942852 Category : Science Languages : en Pages : 492
Book Description
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.
Author: Mouldy Sioud Publisher: Springer Science & Business Media ISBN: 1592597467 Category : Science Languages : en Pages : 623
Book Description
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.
Author: Robert J Scarborough Publisher: Humana ISBN: 9781071607183 Category : Medical Languages : en Pages : 304
Book Description
This volume provides protocols designed to study the function and the structure of diverse ribozymes. Chapters guide readers through different techniques to identify and characterize new ribozymes and methods to use ribozymes to alter the function of CRISPR-based guide RNAs, AgoshRNAs and aptamers or to study RNA capping and long non-coding RNAs. 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 tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Ribozymes: Methods and Protocols aims to help in accelerating ribozyme research and inspiring others to develop new methods to study ribozyme structure and function.
Author: Daniel A Lafontaine Publisher: Humana Press ISBN: 9781627037310 Category : Medical Languages : en Pages : 278
Book Description
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.
Author: John J. Rossi Publisher: Taylor & Francis ISBN: 9781898486176 Category : Science Languages : en Pages : 294
Book Description
In this definitive guide to intracellular ribozyme applications, researchers from both industry and academia review the most recent pharmaceutical, therapeutic, and biotechnological applications of ribozymes. Leading scientists from around the world present the latest principles on the application of ribozyme technology to the inhibition of gene expression in living cells. The authors are experts in the field and describe, in detail, protocols for monitoring ribozyme activity, for expressing ribozymes from DNA vectors, for delivering ribozymes to cells, and for in vitro selection of improved ribozymes. The book provides a forward-looking view of ribozyme technology for alteration of gene expression in plants and in treatment of retroviral and genetic diseases in man.
Author: Ian Gibson Publisher: John Wiley & Sons ISBN: 3527612521 Category : Science Languages : en Pages : 92
Book Description
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.
Author: David Malcolm James Lilley Publisher: Royal Society of Chemistry ISBN: 0854042539 Category : Science Languages : en Pages : 339
Book Description
Takes the reader through the origins of catalysis in RNA and necessarily includes significant discussion of structure and folding. The main focus of the book concerns chemical mechanism with extensive comment on how, despite the importance of RNA catalysis in the cell, its origins are still poorly understood and often controversial. The reader is given an outline of the important role of RNA catalysis in many aspects of cell function, including RNA processing and translation.
Author: Kevin J. Scanlon Publisher: Humana ISBN: 9781489943439 Category : Science Languages : en Pages : 462
Book Description
The purpose of Applications Therapeutic of Ribozymes is to provide an overview of the utility of ribozymes to selectively inhibit the expression of RNA. The ribozyme applications appearing in this book should benefit not only those experienced in ribozymes, but also those applying this ribozyme technology for the first time. It is hoped that a better understanding of the therapeutic application ofribozymes will have a significant impact on human disease in the near future. The field ofribozyme biochemistry has come a long way since the initial observation that RNA is capable of catalysis, initially in cis during processing of larger molecules and ultimately the use of ribozymes in trans to achieve cleavage of target sequence. The fundamental observation that hammerhead (and subsequently hairpin) ribozymes could be designed to cleave a target messenger RNA, and the clarification of sequence restrictions in both the ribozyme and the target RNA, have paved the ways for the use of ribozymes as a tool to manipulate gene expression at the molecular level. This had pre- ously been a domain of antisense oligonucleotides and triplex DNA. Sub- quent studies have explored the myriad biological systems in which ribozyme-mediated inhibition of genes involved in various cellular processes may be used to uncouple important signaling pathways or to reverse the p- notypic expression of a pathologic process.
Author: Mouldy Sioud Publisher: Springer Science & Business Media ISBN: 1597451657 Category : Medical Languages : en Pages : 360
Book Description
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.
Author: Benoît Masquida Publisher: John Wiley & Sons ISBN: 1786309777 Category : Science Languages : en Pages : 196
Book Description
Behind the neologism “ribozymes” lies a family of fascinating molecules, ribo-enzymes, which have been relatively little studied. These catalytically active RNAs are found in all strata of life, from viruses to the human genome. At the end of the 1970s, the discovery of a catalytic RNA nestled in an intron, followed by another involved in the maturation of transfer RNAs, led to the discovery of new ribozymes and the transition from a strictly “proteocentric” vision, inherited from the dogma of molecular biology, to a more “nucleocentric” one. Since then, a variety of ribozymes have been identified in genomes, where their functions often remain mysterious. Looking at Ribozymes traces the discovery of these molecules and presents a picture of their functional diversity, catalytic mechanisms and distribution within the tree of life.
Author: Philip C. Turner
Author: Philip C. Turner
Author: Mouldy Sioud
Author: Robert J Scarborough
Author: Daniel A Lafontaine
Author: John J. Rossi
Author: Ian Gibson
Author: David Malcolm James Lilley
Author: Kevin J. Scanlon
Author: Mouldy Sioud
Author: Benoît Masquida