400 pages | 4 Color Illus. | 68 B/W Illus.
Maximizing the potential of RNA interference in functional genomics - as well as in the development of therapeutics - continues to be at the forefront of biomedical research. Unlike journal articles, Gene Silencing by RNA Interference: Technology and Application combines essential background to the RNAi field with practical techniques designed by renowned researchers to provide the most diverse and in-depth examination of the subject yet.
This book describes methods and protocols for gene silencing and RNA interference. Each chapter provides necessary background to the subject and then gives detailed methods in easy-to-follow steps, along with troubleshooting hints and tips. Following a general and historical introduction, chapters two through eight focus on technical details of the various methods of siRNA design, chemical and enzymatic production, plasmid- and virus-mediated intracellular expression, and other RNAi tools. Chapter nine discusses the exogenous delivery of siRNAs into cells, and the final chapters of the book detail the application of RNAi to dissect gene function in a number of biological systems, including cell-free systems, cultured cells, and whole organisms.
From the design and production of RNAi tools to their applications, Gene Silencing by RNA Interference: Technology and Application is the first to present the entire spectrum of activity, design, production, and delivery of RNAi reagents, providing a welcome guide for both academic and commercial use.
Gene silencing by RNA interference and the role of small interfering RNAs; N.J. Caplen, National Institutes of Health, Maryland, USA
Basics of siRNA design and chemical synthesis; Q. Boese, A. Reynolds, and W.S. Marshall, Dharmacon, Inc. Colorado, USA
Oligonucleotide scanning arrays in the design of small interfering RNAs; V.M. Macaulay and M. Sohail, Weatherall Institute of Molecular Medicine and University of Oxford, England, UK
siRNA production by in vitro transcription; O. Donzé, P. Dudek, and D. Picard, Université de Genève and Apotech Corporation, Switzerland
Production of siRNAs with the application of deoxyribozymes; G. Doran and M. Sohail, University of Oxford, England, UK
Production of siRNA in vitro by enzymatic digestion of double-stranded RNA; F. Buchholz, D. Drechsel, M. Ruer, and R. Kittler, Max Planck Institute, Dresden, Germany
Plasmid-mediated intracellular expression of siRNAs; R. Wadhwa, M. Sano, M. Miyagishi, and K. Taira, National Institute of Advanced Industrial Science and Technology, Tsukuba, and University of Tokyo, Japan
Lentiviral vector-mediated delivery of si/shRNA, D.S. An, X-F. Qin, Vincent Auyeung, D. Baltimore, and I.S.Y. Chen, University of California-Los Angeles, UT MD Anderson Cancer Center in Texas, and California Institute of Technology, Pasadena, USA
Exogenous siRNA delivery: protocols for optimizing delivery to cells; A.J. Hollins, S.P. Fox, and S. Akhtar, Cardiff University, Wales, UK
RNAi in Drosophila cell cultures, M. Bettencourt-Dias, R. Sinka, L. Frenz, and D. Glover, University of Cambridge. and Cyclacel, Ltd., England, UK
RNAi in Caenorhabditis elegans; F. Simmer and R.H.A. Plasterk, Center for Biomedical Genetics, Utrecht, The Netherlands
Delivery of RNAi reagents in C. elegans by microinjection; P. Appleford and A. Woollard, University of Oxford, England, UK
RNAi in Drosophila embryos; R.W. Williams, University of California, Berkeley, USA
RNAi in Xenopus laevis oocytes; E. Gordon and G.W. Abbott, Cornell University, New York, USA
RNAi in Xenopus embryos; K.H. Kok and D-Y. Jin, University of Hong Kong, People's Republic of China
An RNAi-based genomic library for forward genetics in the African Trypanosome; J.C. Morris, Z. Wang, S.A. Motyka, M.E. Drew, and P.T. Englund, Clemson University, South Carolina, Massachusetts Institute of Technology, Massachusetts, Johns Hopkins University, Maryland, and Washington University, Missouri, USA
RNAi in the malaria parasite Plasmodium; A. Brüning-Richardson and G.A. McConkey, University of Leeds, England, UK
Silencing the expression of cysteine proteases (falcipains) in Plasmodium falciparum using RNA interference approaches; P.V.N. Dasaradhi, A. Mohmmed, M.J. Hossain, V.S. Chauhan, and P. Malhotra, International Center for Genetic Engineering and Biotechnology, New Delhi, India
RNA in transgenic animal models; H. Hasuwa & M. Okabe, Osaka University, Japan
RNA interference in mouse model, E. Song, P. Stern, D. Palliser, Luk Van Parijs and J. Lieberman, Harvard University, Massachusetts, USA
Virus-induced RNA silencing in plants; M. Figlerowicz, M. Barciszewska-Pacak, B. Grabowska, J. Wojciechowicz, and A. Pacak, Adam Mickiewicz University, Poznan, University of Warmia and Mazury, Olsztyn, and Polish Academy of Sciences, Poznan, Poland
RNA silencing in plants - biolistic delivery of RNAi reagents; U. Klahre and F. Meins, Novartis Research Foundation, Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland
Agro-infiltration: a versatile tool for RNAi studies in plants; D. Silhavy, Agricultural Biotechnology Center, Godollo, Hungary