Laboratory Protocols for Conditional Gene Targeting PDF Download
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Author: Raul M. Torres
Publisher:
ISBN:
Category :
Languages : en
Pages :
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Book Description
Author: Raul M. Torres
Publisher:
ISBN:
Category :
Languages : en
Pages :
Get Book Here
Book Description
Author: Raul M. Torres
Publisher:
ISBN:
Category :
Languages : en
Pages : 167
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Book Description
Author: Ralf Kühn
Publisher: Humana Press
ISBN: 9781607613060
Category : Science
Languages : en
Pages : 516
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Book Description
Following the completion of the mouse and human genome sequences, a major challengeisthefunctionalcharacterizationofeverymammaliangeneandthedeciph- ing of their molecular interaction network. The mouse offers many advantages for the use of genetics to study human biology and disease, unmatched among other m- mals. Its development, body plan, physiology, behavior, and diseases have much in common, based on the fact that 99% of the human genes have a mouse ortholog. The investigation of gene function using mouse models is based on many years of tech- logical development. In the two decades since gene targeting in murine embryonic stem (ES) cells was first described by Mario Capecchi and colleagues, more than 3000 predesigned mouse mutants have been developed. To date, a variety of mouse mutagenesis techniques, either gene- or phenotype-driven, are used as systematic approaches. The availability of the genome sequence supports gene-driven approaches such as gene-trap and targeted mutagenesis in ES cells, allowing efficient and precise gene disruption. In combination with the use of site-specific DNA recombinases, in particular the Cre/loxP system, gene disruptioncan be directed to specific cell types in conditionalmousemutants. Furthermore,chemicalandtransposonmutagenesisofthe mouse genome enables us to perform phenotype-driven screens for the unbiased identification of phenotype–genotype correlations involved in models of human d- ease. Over the next several years, the mouse genome will be systematically altered, and the techniques for achieving predesigned manipulations will be constantly developed further and improved. The second edition of Gene Knockout Protocols brings together distinguished c- tributorswithextensiveexperienceinthegenetargetingandmousegeneticsfields.
Author: Jianghui Hou
Publisher: Academic Press
ISBN: 0128226439
Category : Science
Languages : en
Pages : 422
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Book Description
A Laboratory Guide to the Tight Junction offers broad coverage of the unique methods required to investigate its characteristics. The methods are described in detail, including its biochemical and biophysical principles, step-by-step process, data analysis, troubleshooting, and optimization. The coverage includes various cell, tissue, and animal models. Chapter 1 provides the foundations of cell biology of tight junction. Chapter 2 covers the Biochemical approaches for paracellular channels and is followed by chapter 3 providing the Biophysical approaches. Chapter 4 describes and discusses Histological approaches for tissue fixation and preparation. Chapter 5 discusses Light microscopy, while chapter 6 presents Electron microscopic approaches. Chapter 7 covers Transgenic manipulation in cell cultures, including DNA and siRNA, Mutagenesis, and viral infection. Chapter 8 covers transgenic manipulation in mice, including: Knockout, Knockin, siRNA knockdown, GFP/LacZ reporter, and overexpression. The final chapter discusses the future developments of new approaches for tight junction research. Researchers and advanced students in bioscience working on topics of cell junction, ion channel and membrane protein will benefit from the described methods. Clinicians and pathologists interested in tissue barrier diseases will also benefit from the biochemical and biophysical characterization of tight junctions in organ systems, and their connection to human diseases. Provides consistent and detailed research methods Covers various cell, tissue and animal models Includes step-by-step guidance from beginner to sophisticated levels
Author: Alexandra Joyner
Publisher: OUP Oxford
ISBN: 0191566020
Category : Science
Languages : en
Pages : 312
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Book Description
Since the publication of the first edition of Gene Targeting: A Practical Approach in 1993 there have been many advances in gene targeting and this new edition has been thoroughly updated and rewritten to include all the major new techniques. It provides not only tried-and-tested practical protocols but detailed guidance on their use and applications. As with the previous edition Gene Targeting: A Practical Approach 2e concentrates on gene targeting in mouse ES cells, but the techniques described can be easily adapted to applications in tissue culture including those for human cells. The first chapter covers the design of gene targeting vectors for mammalian cells and describes how to distinguish random integrations from homologous recombination. It is followed by a chapter on extending conventional gene targeting manipulations by using site-specific recombination using the Cre-loxP and Flp-FRT systems to produce 'clean' germline mutations and conditionally (in)activating genes. Chapter 3 describes methods for introducing DNA into ES cells for homologous recombination, selection and screening procedures for identifying and recovering targeted cell clones, and a simple method for establishing new ES cell lines. Chapter 4 discusses the pros and cons or aggregation versus blastocyst injection to create chimeras, focusing on the technical aspects of generating aggregation chimeras and then describes some of the uses of chimeras. The next topic covered is gene trap strategies; the structure, components, design, and modification of GT vectors, the various types of GT screens, and the molecular analysis of GT integrations. The final chapter explains the use of classical genetics in gene targeting and phenotype interpretation to create mutations and elucidate gene functions. Gene Targeting: A Practical Approach 2e will therefore be of great value to all researchers studying gene function.
Author: Francesca Storici
Publisher: Humana Press
ISBN: 9781627037624
Category : Medical
Languages : en
Pages : 463
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Book Description
Gene correction is a technology that gives us the tools for both repairing and mutating DNA, for discovering gene functions and for engineering new genetic variants. Gene Correction: Methods and Protocols provides a user friendly, detailed and up-to-date collection of strategies and methodologies utilized for generating specific sequence changes in the DNA of cells in the laboratory, while also tackling the major problems that the field of gene correction faces. This volume brings together many experts in the field of gene correction to disclose a wide and varied array of specific gene correction protocols for engineering mutations in DNA, for delivering correcting DNA to target cells, and for improving the accuracy and safety of the gene correction process. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Gene Correction: Methods and Protocols seeks to serve scientists of all backgrounds interested in the area of gene targeting/recombination/therapy.
Author: Marten H. Hofker
Publisher: Springer Science & Business Media
ISBN: 1592593402
Category : Science
Languages : en
Pages : 382
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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.
Author: Hans Hedrich
Publisher: Academic Press
ISBN: 0123820081
Category : Medical
Languages : en
Pages : 869
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Book Description
"Mice have long been recognized as a valuable tool for investigating the genetic and physiological bases of human diseases such as diabetes, infectious disease, cancer, heart disease, and a wide array of neurological disorders. With the advent of transgenic and other genetic engineering technologies, the versatility and usefulness of the mouse as a model in biomedical research has soared. As a result, mouse colonies everywhere are expanding, and scientists who previously focused on other models are turning their attention to the mouse. Revised to reflect advances since the first edition, The Laboratory Mouse, Second Edition continues to be the most accessible reference on the biology and care of the laboratory mouse. This guide presents basic information and common procedures in detail to provide a quick reference source for investigators, technicians, and caretakers in the humane care and use of the mouse in the laboratory setting. Expanded, updated, and now in color, this new edition includes coverage of the biological features, husbandry, management, veterinary care, experimental methodology, and resources applying specifically to the mouse"--Provided by publisher.
Author: Shirley Pease
Publisher: Springer Science & Business Media
ISBN: 3540284893
Category : Science
Languages : en
Pages : 300
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Book Description
Mammalian and Avian Transgenesis presents a collection of novel methods for the production of a wide range of transgenic animals. The manual focuses largely on mice, but also contains protocols for successful transgenesis in rats, cows, pigs and birds. The manual provides detailed, step-by-step protocols covering all aspects of the production of transgenic animals, including the use of lentiviral vectors in gene transfer, intracytoplasmic sperm injection, nuclear transfer, large insert transgenesis, conditional gene expression systems, the use of reporter genes in transgenesis and transgenesis in large animals and birds. The text is supplemented by superb color photos. While the focus is on newly established techniques, the fundamental methods of transgenesis are also covered for those new to the field. Thus this manual is perfectly suited for those wishing to adopt new technologies in transgenesis.
Author: Alan R. Clarke
Publisher: Springer Science & Business Media
ISBN: 1592591787
Category : Science
Languages : en
Pages : 346
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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.
