Antisense Technologies

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.

Readership: Graduate and post-doctoral researchers and academic staff.

Edited by Conrad Lichtenstein, Professor, School of Biological Sciences, Queen Mary and Westfield College, London, and Wolfgang Nellen, Abteilung Genetik FB19, Universitat Kassel, Kassel, Germany

"Genetic researchers will find ANTISENSE TECHNOLOGIES a most valuable source of information." - Library of Science, Winter 1998

"providing clear and precisely described protocols which cover a broad range of applications.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 434 (1998) 442-446

"'...this volume reaches the standard we have come to expect from the Practical Approach series. The standard of presentation reflects the expertise of the authors and the protocol are described clearly with useful practical notes.'" - Glenn Matthews, SGM Quarterly Nov. 1998.

1: Nucleic acid structures
2: Evaluation of antisense effects
3: Selecting, preparing, and handling antisense oligodeoxyribonucleotides
4: In vitro RNAs
5: Catalytic antisense RNA based on hammerhead ribozymes
6: 2-5 A-Antisense chimaeras for targeted degradation of RNA
7: In vitro applications of antisense RNA and DNA
8: Antisense applications in Dictyostelium: a lower eukaryotic model system
9: Applications of antisense technology in plants
10: Antisense molecules and ribozymes: medical applications
11: Non-antisense effects of oligodeoxynucleotides
12: Antisense rescue
13: Shotgun antisense mutants
14: Detection of sense-antisense duplexes by structure-specific anti-RNA antibodies