This hands on guide to RNA interference brings the power of targeted gene silencing to any laboratory with the basic equipment for handling nucleic acids. In easy to follow, step by step protocols you will learn: how RNAi works in worms, flies and mammals; how to design the most efficient RNAi constructs; how to achieve transient, stable and conditional RNAi in cell cultures; how to determine the efficiency of an RNAi experiment; and how to use RNAi for gene therapy. All the protocols have been thoroughly tested in the author's own laboratory, and she provides examples of successful experiments and troubleshooting hints to help in establishing your own successful RNAi experiments. It also includes a list of suppliers for RNAi reagents and equipment as well as a glossary of terms.
Ute Schepers studied Chemistry at the Kekule-Institut fur Organische Chemie und Biochemie, University of Bonn (Germany), where she completed her PhD thesis in the laboratory of Prof. Dr. K. Sandhoff in 1997. She then moved to Boston for a post-doctoral fellowship in the laboratory of Tomas Kirchhausen at the Department of Cell Biology, Harvard Medical School. Since her return to Bonn in 2000 she is pursuing her own research projects using a combination of chemical and genetic methods for the further development of RNA interference in mammals. One of her main efforts is the chemical synthesis of novel molecular transporters for therapeutical RNAi application.
Preface. 1 Introduction: RNA interference, the "Breakthrough of the Year 2002". 2 RNAi in Caenorhabditis elegans. 3 RNAi in Drosophila. 4 RNAi in Mammals. Appendix 1: Abbreviations. Appendix 2: List of Protocols. Appendix 3: Suppliers of RNAi-related Chemicals and Probes. Appendix 4: Glossary of Terms. Subject Index.