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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. Also includes a list of suppliers for RNAi reagents and equipment as well as a glossary of terms.
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
RNA interference (RNAi) is a widely used technology for gene silencing and has become a key tool in a myriad of research and lead discoveries. In recent years, the mechanism of RNAi agents has been well investigated, and the technique has been optimized for better effectiveness and safety. On the other hand, the clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas9/gRNA system is a recent, novel, targeted genome-editing technique derived from the bacterial immune system. Recent advances in gene-editing research and technologies have enabled the CRISPR Cas9 system to become a popular tool for sequence-specific gene editing to correct and modify eukaryotic systems. In this book, we will focus on the mechanisms, applications, regulations (their pros and cons), and various ways in which RNAi-based methods and CRIPSR-Cas9 technology have stimulated the modulation of gene expression, thereby making them a promising therapeutic tool to treat and prevent complex diseases and disorders.
A second edition of the classic handbook has become a standard in the Drosophila field. This edition is expanded to include topics in which classical genetic strategies have been augmented with new molecular tools. Included are such new techniques as homologous recombination, RNAi, new mapping techniques, and new mosaic marking techniques.
New Findings Revolutionize Concepts of Gene FunctionEndogenous small RNAs have been found in various organisms, including humans, mice, flies, worms, fungi, and bacteria. Furthermore, it's been shown that microRNAs acting as cellular rheostats have the ability to modulate gene expression. In higher eukaryotes, microRNAs may regulate as much as 50 p
This book offers a range of environmentally benign molecular mechanisms which are safer alternative strategies for effective insect pest management. In modern era of biotechnology, there has been much advancement in the field of molecular biology, where many more techniques have evolved which can be helpful in the field of pest management too. Plant resistance, development of transgenic plants, and many more techniques are being considered the panacea to pest problems. On the other hand, there are wide spread concerns of the safety of biotechnological interventions with nontarget organisms including humans. While the world stands divided on the ethical issues of these approaches and the many safety concerns, scientists believe that well thought of biotechnological interventions are probably the only safest ways possible for reducing pest attacks on crops. It explores various techniques and aspects related to molecular pathways for crop pest control. This book is a useful resource for postgraduate students and researchers of agriculture sciences, plant pathology and plant physiology. It is also useful for policy planners in agriculture.
This volume provides readers with a comprehensive review of various gene silencing methodologies and its applications. It provides a historical overview of gene silencing mechanisms in plants, vectors, and strategies available for plant gene silencing, practical applications of gene silencing, bioinformatics tools, and other resources. In addition to these review chapters, this book includes methodology for virus-induced gene silencing (VIGS) in various different plant species, understanding plant stress responses using VIGS, miRNA identification, DNA interference, host-induced gene silencing, use of artificial miRNAs for gene silencing, high throughput RNAi, and more. Written in the highly successful Methods of Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Thorough and practical, Plant Gene Silencing: Methods and Protocols seeks to aid scientists to further understand functional relevance of target genes using gene silencing methods and use these technologies in commercial plant varieties.
The enormous potential of siRNA as a therapeutic has led to an explosion of interest from the scientific community. There has been intense interest from Big Pharma to capitalise on this new technology but the fact remains that delivery is a key determinant in realizing the full clinical potential of RNA interference. There is an urgent need for better delivery methods to take this technology forward. This book addresses the role of different RNAi molecules in cellular processes as rational for diagnostic and therapeutic approaches. This book will cover RNAi therapeutic design to optimize siRNA potency and reduce off-target effects and current delivery technologies to overcome both intracellular and extracellular barriers. The reader will gain an insight into RNA interference from the cellular mechanisms to screening to siRNA design right through to diagnostic and therapeutic applications.