Antisense technology is a powerful procedure that permits the controlled silencing of a specific gene for investigations of mRNA and protein function. This valuable text provides proven step-by-step protocols for antisense techniques in a range of different organisms and cell culture systems. In addition it discusses the potential benefits and problems for various antisense methods which complement gene knock-out experiments. The book includes antisense techniques such as: analysis of nucleic acid structures; measurement and evaluation of antisense effects; selection, preparation and the use of antisense oligonucleotides; in vitro RNA transcription; construction strategies, testing and optimization of catalytic antisense RNAs based on hammerhead ribozymes; synthesis and evaluation of 2-5 A- antisense chimeras for targeted degradation of RNA. The application of these technologies are then described, with chapters on antisense techniques in IDictyostelium, plants and the medical uses and benefits of antisense sense technology both in vitro and in vivo. Finally, non-antisense effects of oligonucleotides, "anti-sense rescue", shotgun antisense mutagenesis, and in vivo, in vitro and in situ detection of sense: antisense duplexes by structure-specific anti-RNA antibodies are discussed. The numerous hints and tips for success and advice on trouble shooting, make this authoritative text invaluable to all those researchers who work on gene expression, RNA transcription, or protein function.
"The antisense strategy represents an attractive approach to control gene expression by restoring normal regulatory functions or to investigate the role of specific mRNA and protein. Nevertheless, in order to achieve successful results, skilful ability in managing this technology and optimization of the methodology are required. This book fulfils such expectations by providing clear and precisely described protocols . . . Antisense Technology offers potential for therapy and for the study of gene roles through the development of specific agents that can modulate the function of the genes or their proteins. The experience gained from experimental and clinical trials in the antisense area has been concentrated in this book which provides updated data and methodologies that may be extremely useful for everybody who is already familiar with antisense oligonucleotides and for those investigators who are approaching this field in continuous progress for the first time."--FEBS Letters
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