Gene Silencing By Rna Interference

RNAi technology is used for large-scale screens that systematically shut down each gene in the cell, which can help identify the components necessary for a particular cellular process or an event such as cell division. Exploitation of the pathway is also a promising tool in biotechnology and medicine. Introducing new technology in the study of RNA

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.

This book focuses on an emerging, central issue in molecular genetics and the development of eukaryotes: the control of gene expression by small species of RNA. As an exciting new field of endeavor, it is the first book by a single author to deal comprehensively with RNA silencing. The book provides the historical background of the field preceding the seminal work by Fire and associates in 1998 on the impact of small double-stranded RNA on the expression of nematode genes, which is considered the beginning of RNA silencing research. RNA silencing is described in a wide range of plants and animals including protozoa, simple metazoa, insects, non-mammalian vertebrates, and mammals. In each case the experimental results are provided with the accompanying background and with illustrations. There is also an appendix on the prospective use of RNA silencing in gene therapy, which is intended as a guide for investigators wishing to explore this possibility.

In the past three years, the use of double-stranded RNA to silence gene activity has become widely and rapidly adopted. RNA interference is highly specific and remarkably potent, and it acts on cells and tissues far removed from the site of introduction. The principles behind RNAi are just being uncovered, but this laboratory technique has been applied effectively in a wide variety of animal and plant species. Variations on RNAi are revolutionizing many approaches to experimental biology, complementing traditional genetic technologies with a quicker and less expensive way of mimicking the effects of mutations both in cell cultures and in living animals. Recent advances in the use of RNAi to engineer heritable silencing in mammals, to alter stem cells for organ reconstitution, and to alter the course of disease in model systems indicate that RNAi may have a future in disease therapy. Written by pioneers in this new field and edited by Gregory Hannon, one of its leading figures, RNAi: A Guide to Gene Silencing presents the principles of RNAi and reliable protocols for its laboratory use in Caenorhabditis elegans, Drosophila, plants, avian embryos, mammalian cells, mouse oocytes, and more. This important and unique book is an essential laboratory resource for scientists studying gene regulation and for all experimental biologists interested in the emerging practical applications of RNAi.

A collection of readily reproducible methods for the design, preparation, and use of RNAs for silencing gene expression in cells and organisms. The techniques range widely and include methods addressing the biochemical aspects of the silencing machinery, RNA silencing in non-mammalian organisms, and the in vivo delivery of siRNAs and silencing vectors. There are also techniques for designing, preparing, and using RNAs to silence gene expression, for fine-tuning regulation by targeting specific isoforms of a given gene, and for the study and use of microRNAs. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

RNA Interference (RNAi) technology has rapidly become one of the key methods used in functional genomics. RNAi is used to block the expression of genes and create phenotypes that can potentially yield clues about the function of these genes. In the postgenomic era, the elucidation of the physiological function of genes has become the rate-limiting step in the quest to develop 'gene-based drugs' and RNAi could potentially play a pivotal role in the validation of such novel drugs. In this overview, the basic concepts and applications of RNAi biology are discussed. Leading experts from both academia and industry have contributed to this invaluable reference. The volume is forwarded by Andrew Fire, one of the winners of the 2006 Nobel Prize for the discovery of RNA Interference.

An in-depth treatment of cutting-edge work being done internationally to develop new techniques in crop nutritional quality improvement Phytonutritional Improvement of Crops explores recent advances in biotechnological methods for the nutritional enrichment of food crops. Featuring contributions from an international group of experts in the field, it provides cutting-edge information on techniques of immense importance to academic, professional and commercial operations. World population is now estimated to be 7.5 billion people, with an annual growth rate of nearly 1.5%. Clearly, the need to enhance not only the quantity of food produced but its quality has never been greater, especially among less developed nations. Genetic manipulation offers the best prospect for achieving that goal. As many fruit crops provide proven health benefits, research efforts need to be focused on improving the nutritional qualities of fruits and vegetables through increased synthesis of lycopene and beta carotene, anthocyanins and some phenolics known to be strong antioxidants. Despite tremendous growth in the area occurring over the past several decades, the work has only just begun. This book represents an effort to address the urgent need to promote those efforts and to mobilise the tools of biotechnical and genetic engineering of the major food crops. Topics covered include: New applications of RNA-interference and virus induced gene silencing (VIGS) for nutritional genomics in crop plants Biotechnological techniques for enhancing carotenoid in crops and their implications for both human health and sustainable development Progress being made in the enrichment and metabolic profiling of diverse carotenoids in a range of fruit crops, including tomatoes, sweet potatoes and tropical fruits Biotechnologies for boosting the phytonutritional values of key crops, including grapes and sweet potatoes Recent progress in the development of transgenic rice engineered to massively accumulate flavonoids in-seed Phytonutritional Improvement of Crops is an important text/reference that belongs in all universities and research establishments where agriculture, horticulture, biological sciences, and food science and technology are studied, taught and applied.

The critically acclaimed laboratory standard, Methods in Enzymology, is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. The series contains much material still relevant today - truly an essential publication for researchers in all fields of life sciences. RNA Interference will cover RNAi in non-vertebrates (plants, C. elegans, drosophila, and S. pombe), and Mammalian systems (human and non-human cells). This volume discusses extensive methodology related to delivery methods high throughput strategies and prospects as a human therapy agent. * One of the most highly respected publications in the field of biochemistry since 1955 * Frequently consulted, and praised by researchers and reviewers alike * Truly an essential publication for anyone in any field of the life sciences