Transgenesis Techniques


Book Description




Advanced Protocols for Animal Transgenesis


Book Description

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.




Safety of Genetically Engineered Foods


Book Description

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.




Transgenesis Techniques


Book Description

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.




Transgenic Mouse Methods and Protocols


Book Description

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.




Animal Transgenesis and Cloning


Book Description

Designed to be a concise, balanced introduction to this dynamic subject, covering key issues and current techniques currently used in animal transgenesis and cloning. Provides the essentials of the subject, from the molecular basis of gene structure and function to therapeutic gene cloning in humans. Social and ethical implications of this important area of research are also considered.




Transgenesis Techniques


Book Description

One of the major challenges currently facing the scientific community is to understand the function of the multitude of protein-coding genes that were revealed when the human genome was fully sequenced. In Transgenesis Techniques: Principles and Protocols, Third Edition, experts in the field fully update and expand upon the previous edition in order to detail the transgenic techniques currently used to modify the genome. The volume explores classic procedures to genetically modify mice and other model organisms, as well as cutting-edge practices involving microinjection, site-specific recombination systems, cryopreservation, and many other topics. Written in the highly successful Methods in Molecular BiologyTM series format, the chapters include brief introductions to their subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls. Comprehensive and state-of-the-art, Transgenesis Techniques: Principles and Protocols, Third Edition is a comprehensive practical guide to the generation of transgenic animals and an invaluable source of information in any lab currently involved in transgenic techniques.




Transgenic Animals


Book Description

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.




Rabbit Biotechnology


Book Description

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.




Nuclear Transfer Protocols


Book Description

Nuclear Transfer Protocols: Cell Reprogramming and Transgenesis is a comprehensive review of nuclear transfer technology in vertebrates, aimed at reprogramming differentiated nuclei and effecting targeted gene transfer. The emphasis here is on providing readily reproducible techniques for the gene- tion of cloned embryos and animals in a number of key research and commercially important vertebrates. Additional chapters provide alternative cutting-edge methods for nuclear transfer, such as zona-free nuclear transfer and serial nuclear transfer. Of immense practical benefit are descriptions of procedures associated with cloning, such as in vitro maturation of oocytes, activation and culture of cloned embryos, maintenance of pregnancy, and neonatal care of clones. Nuclear Transfer Protocols: Cell Reprogramming and Transgenesis also provides an understanding of the factors involved in nuclear reprogramming, which is imperative for the success of cloning. A section dealing with such cloning-related issues as aging and normality of clones is also included making this an essential comprehensive handbook for research and commercial labo- tories involved in, or intending to work on, nuclear transfer. The volume will prove beneficial to molecular biologists, stem cell biologists, clinicians, biotechnologists, students, veterinarians, and animal care technicians involved with reprogramming, nuclear transfer, and transgenesis.