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.
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.
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.
This detailed book explores how microinjection will be used in the foreseeable future, not only for generating animal models for biomedical research but also for changing economically or ecologically important species that can broadly impact our society in general. The opening half of the book focuses on methods for generating mouse models, as they are still the most popular in genome engineering research, while the second half examines gene-editing in a variety of other species, opened up by the developments in ZFN, TALEN, and CRISPR techniques. Written for the highly successful Methods in Molecular Biology series, 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. Authoritative and practical, Microinjection: Methods and Protocols serves as an ideal guide for researchers looking to take advantage of the breakthrough technologies in gene-editing and embryo micromanipulations.
The laboratory mouse is an important model for addressing questions in cancer biology. In recent years, the questions have become more refined, and mouse models are increasingly being used to develop and test cancer therapeutics. Thus, the need for more sophisticated and clinically relevant mouse models has grown, as has the need for innovative tools to analyze and validate them. This laboratory manual provides cutting-edge methods for generating and characterizing mouse models that accurately recapitulate many features of human cancer. The contributors describe strategies for producing genetic models, including transgenic germline models, gene knockouts and knockins, and conditional and inducible systems, as well as models derived using transposon-based insertional mutagenesis, RNA interference, viral-mediated gene delivery, and chemical carcinogens. Tissue recombination, organ reconstitution, and transplantation methods to develop chimeric, allograft, and xenograft models are covered. Approaches to characterize tumor development, progression, and metastasis in these models using state-of-the-art imaging, histopathological, surgical, and other techniques are also included. Other chapters cover the use of mouse models to test and optimize drugs in pre-, co-, and post-clinical trials. An appendix specifically addresses the use of mouse cancer models in translational studies and the integration of mouse and human clinical investigations. This manual is therefore an indispensable laboratory resource for all researchers, from the graduate level upwards, who study cancer and its treatment.
Cultured cells have combined accessibility and the ability to expand a homogeneous cell population from a relatively limited source, thus opening up a wealth of possibilities for researchers. In Mouse Cell Culture: Methods and Protocols, expert researchers provide a number of methods for the culture of a wide range of specific cells and tissues isolated from the key genetic model of the fetal or adult mouse. Including protocols for the explant of fetal tissues and stem cells that allow developmental processes to be followed ex vivo as well as protocols for the culture of isolated cell types that allow for the study of relatively homogeneous cell populations, this volume brings together a selection of the most current methods in order to make them available in one convenient source. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Practical and authoritative, Mouse Cell Culture: Methods and Protocols serves as an immediately applicable springboard for the development of new tissue culture methods in order to advance the study and treatment of human disorders.
Using the most well-studied behavioral analyses of animal subjects to promote a better understanding of the effects of disease and the effects of new therapeutic treatments on human cognition, Methods of Behavior Analysis in Neuroscience provides a reference manual for molecular and cellular research scientists in both academia and the pharmaceutic
Most people have some interest in embryos; this probably results, in part, from their interest in understanding the biological origins of themselves and their offspring and, increasingly, concerns about how environmental change such as pollution might affect human development. Obviously, et- cal considerations preclude experimental studies of human embryos and, c- sequently, the developmental biologist has turned to other species to examine this process. Fortunately, the most significant conclusion to be drawn from the experimental embryology of the last two decades is the manner in which orthologous or closely related molecules are deployed to mediate similar - velopmental processes in both vertebrates and invertebrates. The molecular mechanisms regulating processes fundamental to most animals, such as axial patterning or axon guidance, are frequently conserved during evolution. (It is now widely believed that the differences between phyla and classes are the result of new genes, arising mostly by duplication and divergence of extant sequences, regulating the appearance of derived characters. ) Other vertebrates are obviously most likely to use the same devel- mental mechanisms as humans and, within the vertebrate subphylum, the - parent degree of conservation of developmental mechanism is considerable. It has long been recognized that particular vertebrate species offer either d- tinct advantages in investigating particular stages of development or are - pecially amenable to particular manipulations. No single animal can provide all the answers because not all types of experiments can be carried out on a single species.
