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The past decade has witnessed a spectacular explosion in both the devel- ment and use of transgenic technologies. Not only have these been used to aid our fundamental understanding of biologic mechanisms, but they have also faci- tated the development of a range of disease models that are now truly beginning to impact upon our approach to human disease. Some of the most exciting model systems relate to neurodegenerative disease and cancer, where the availability of appropriate models is at last allowing radically new therapies to be developed and tested. This latter point is of particular significance given the current concerns of the wider public over both the use of animal models and the merits of using genetically modified organisms. Arguably, advances of the greatest significance have been made using mammalian systems—driven by the advent of embryonic stem-cell–based strategies and, more recently, by cloning through nuclear transfer. For this reason, this new edition of Transgenesis Techniques focuses much more heavily on manipulation of the mammalian genome, both in the general discussions and in the provision of specific protocols.
Assists policymakers in evaluating the appropriate scientific methods for detecting unintended changes in food and assessing the potential for adverse health effects from genetically modified products. In this book, the committee recommended that greater scrutiny should be given to foods containing new compounds or unusual amounts of naturally occurring substances, regardless of the method used to create them. The book offers a framework to guide federal agencies in selecting the route of safety assessment. It identifies and recommends several pre- and post-market approaches to guide the assessment of unintended compositional changes that could result from genetically modified foods and research avenues to fill the knowledge gaps.
This laboratory manual, published in cooperation with the International Society for Transgenic Technology (ISTT), provides almost all current methods that can be applied to the creation and analysis of genetically modified animals. The chapters have been contributed by leading scientists who are actively using the technology in their laboratories. Based on their first-hand experience the authors also provide helpful notes and troubleshooting sections. Topics range from standard techniques, such as pronuclear microinjection of DNA, to more sophisticated and modern methods, such as the derivation and establishment of embryonic stem (ES) cell lines, with defined inhibitors in cell culture medium. In addition, related topics with relevance to the field are addressed, including global web-based resources, legal issues, colony management, shipment of mice and embryos, and the three R’s: refinement, reduction and replacement.
Marten Hofker and Jan van Deursen have assembled a multidisciplinary collection of readily reproducible methods for working with mice, and particularlyfor generating mouse models that will enable us to better understand gene function. Described in step-by-step detail by highly experienced investigators, these proven techniques include new methods for conditional, induced knockout, and transgenic mice, as well as for working with mice in such important research areas as immunology, cancer, and atherosclerosis. Such alternative strategies as random mutagenesis and viral gene transduction for studying gene function in the mouse are also presented.
During the past 20 years, transgenesis has become a popular technique and a crucial tool for molecular geneticists and biologists. Transgene expression is now better-controlled and even specifically inducible by exogenous factors. While these techniques have quite significantly transformed the experimental approaches taken by biologists, the applications are more limited than expected and concerns have arisen regarding biosafety as well as physiological, social, and philosophical issues. Transgenic Animals: Generation and Use contains articles on the techniques used to generate transgenic animals and a section on the preparation of vectors for the optimally controlled expression of transgenes. It also examines the use of transgenic animals in the study of gene function and human diseases, the preparation of recombinant proteins and organs for pharmaceutical and medical use, and the improvement of genetic characteristics of farm animals. Finally, it discusses more recent problems generated by transgenic animals including conservation of transgenic lines, specific database patenting, biosafety, and bioethics. Drawn from both academia and industry, the contributors to this monograph present in one concise volume all the relevant information on the different aspects of transgenesis. This book can be used as both a reference book and a textbook for specialized university courses and will be of interest to everyone involved in basic research in animal biology, molecular genetics, animal biotechnology, pharmaceutical science, and medicine.
"The book...is, in fact, a short text on the many practical problems...associated with translating the explosion in basic biotechnological research into the next Green Revolution," explains Economic Botany. The book is "a concise and accurate narrative, that also manages to be interesting and personal...a splendid little book." Biotechnology states, "Because of the clarity with which it is written, this thin volume makes a major contribution to improving public understanding of genetic engineering's potential for enlarging the world's food supply...and can be profitably read by practically anyone interested in application of molecular biology to improvement of productivity in agriculture."
Louis-Marie Houdebine and Jianglin Fan The study of biological functions of proteins and their possible roles in the pathogenesis of human diseases requires more and more relevant animal m- els. Although mice including genetically modified mice offer many possibilities, other non-murine species are absolutely required in some circumstances. Rabbit is one of these species, which has been widely used in biomedical studies. This animal is genetically and physiologically closer to humans including cardiov- cular system and metabolism characteristics. Rabbit is thus more appropriate than mice to study some diseases such as atherosclerosis and lipid metabolism. Because of its larger size, surgery manipulation, bleeding, and turn-over studies are much easier performed in rabbits than in mice. Furthermore, transgenic rabbits can be produced using microinjection and other methods such as lentiviral v- tors. Cloning in rabbits has been proved possible, even though still laborious and time-consuming. Hopefully, functional rabbit ES cell lines will be available in the coming years. Gene deletion or knock-out in rabbits will then become possible.
This volume provides readers with a historical foundation in standard techniques and a comprehensive update on the latest methods used in making gene-modified mice. The chapters in this book cover topics such as pronuclear microinjection in one-cell embryos; embryo transfer surgery; nuclear transfer and cloning; blastocyst microinjection; and cryobanking and recovery of genetically modified mice. Importantly, there are chapters devoted to the latest application of CRISPR technology, as well as the establishment of induced pluripotent stem cells. Written in the highly successful Methods in 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 tips on troubleshooting and avoiding known pitfalls. Comprehensive and authoritative, Transgenic Mouse: Methods and Protocols is a valuable resource for both novice and expert researchers who are interested in learning more about this evolving field.
The aim of Transgenic Plants: Methods and Protocols is to provide a source of information to guide the reader through a wide range of frequently used, broadly applicable, and easily reproducible techniques involved in the gene- tion of transgenic plants. Its step-by-step approach covers a series of methods for genetically transforming plant cells and tissues, and for recovering whole transgenic plants from them. The volume then moves on to the use of sele- able and reporter markers, positive selection, marker elimination after rec- ery of transgenic plants, and the analysis of transgene integration, expression, and localization in the plant genome. Although contributors usually refer to model plants in most chapters, the protocols described herein should be widely applicable to many plant species. The last two sections are devoted to me- ods of risk assessment and to exploring the current and future applications of transgenic technology in agriculture and its social implications in a case study. Transgenic Plants: Methods and Protocols is divided into six major s- tions plus an introduction, comprising 27 chapters. Part I, the Introduction, is a review of the past, present, and perspectives of the transgenic plants, from the discovery of Agrobacterium tumefaciens as a feasible transformation vector, to its use as a tool to study gene expression and function, and the current and possible future applications of this technology in agriculture, industry, and medicine.
Imagine scientists controlling the transmission of certain diseases through the genetic modification of mosquitoes. Eradicating harmful insects without the use of pesticides. Or increasing the fertility of some insects who in turn eat harmful arthropods or even a plant pathogen. Those are just a few of the real-world applications of insect transgen