Download or read book Gene Knockout Protocols written by Martin J. Tymms and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 434 pages. Available in PDF, EPUB and Kindle. Book excerpt: As the major task of sequencing the human genome is near completion and full complement of human genes are catalogued, attention will be focused on the ultimate goal: to understand the normal biological functions of these genes, and how alterations lead to disease states. In this task there is a severe limitation in working with human material, but the mouse has been adopted as the favored animal model because of the available genetic resources and the highly conserved gene conservation linkage organization. In just of ten years since the first gene-targeting experiments were p- formed in embryonic stem (ES) cells and mutations transmitted through the mouse germline, more than a thousand mouse strains have been created. These achievements have been made possible by pioneering work that showed that ES cells derived from preimplantation mouse embryos could be cultured for prolonged periods without differentiation in culture, and that homologous rec- bination between targeting constructs and endogenous DNA occurred at a f- quency sufficient for recombinants to be isolated. In the next few years the mouse genome will be systematically altered, and the techniques for achi- ing manipulations are constantly being streamlined and improved.
Download or read book Gene Knockout Protocols Methods in Molecular Biology written by Martin J. Tymms and published by . This book was released on 2001 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: As the major task of sequencing the human genome nears completion and the full complement of human genes are catalogued, the task of understanding the normal biological functions of genes and how their alteration leads to diseased states becomes more imperative. In Gene Knockout Protocols, highly skilled investigators with extensive experience in gene targeting and mouse genetics describe their best techniques for the design of targeting constructs and for genetic phenotype analysis. These proven methods contain step-by-step instructions, as well as notes on pitfalls to avoid, and emphasize techniques that are relevant to researchers carrying out gene targeting work. These include embryo transplantation, in vitro embryonic stem cell differentiation, creation of aggregation chimeras, mouse pathology, embryo cryopreservation, and transplantation. Issues such as the use of existing mouse mutation resources and the influence of genetic background and epigenetic effects upon phenotype are also covered. State-of-the-art and highly practical, Gene Knockout Protocols not only constitutes an invaluable source of readily reproducible techniques for those just entering the field of gene targeting, but also a key reference for all genetic researchers today.
Download or read book Gene Knockout Protocols written by Ralf Kühn and published by Humana Press. This book was released on 2009-03-27 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book Methods in Molecular Biology Gene knockout protocols written by John M. Walker and published by . This book was released on 1984 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Gene Targeting Protocols written by Eric B. Kmiec and published by . This book was released on 2000 with total page 244 pages. Available in PDF, EPUB and Kindle. Book excerpt: In Gene Targeting Protocols, a panel of innovative investigators present in readily reproducible detail the latest techniques for gene replacement, gene knockout, and gene repair in both animals and plants. These expert researchers review the best gene transfer vehicles (liposomes, PEI, and EPD), discuss the parameters that affect vector uptake, and detail successful methods for gene targeting (Cre-ox, and adeno and adeno-associated viruses). The methods include two fundamentally different approaches to gene targeting- one using a fusion protein to deliver a vector to a virus for possible therapeutic effect, and one entailing an episomal-based Epstein-Barr vector that can modulate the chromatin assembly process, and thus overcome a serious barrier to therapeutic gene targeting. The book also has several techniques for using oligonucleotides in gene targeting, a methodology that may revolutionize the entire field. Innovative and wide ranging, Gene Targeting Protocols offers today's researchers robust and proven gene-targeting techniques that are essential to understanding and regulating biological processes at the genetic level.
Download or read book Genome Editing in Animals written by Izuho Hatada and published by Springer Nature. This book was released on 2023-02-11 with total page 396 pages. Available in PDF, EPUB and Kindle. Book excerpt: This second edition provides new and updated protocols that can be used for generation of knockout animals. Chapters guide the reader through basic protocols for three genome editing technologies, target design tools, and specific protocols for each animal. 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 cutting-edge, Genome Editing in Animals: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field.
Download or read book CRISPR Gene Editing written by Yonglun Luo and published by Humana Press. This book was released on 2019-04-23 with total page 362 pages. Available in PDF, EPUB and Kindle. Book excerpt: This detailed volume guides readers through strategic planning and user-friendly guidelines in order to select the most suitable CRISPR-Cas system and target sites with high activity and specificity. Methods covering CRISPR gRNA design, CRISPR delivery, CRISPR activity quantification (indel quantification), and examples of applying CRISPR gene editing in human pluripotent stem cells, primary cells, gene therapy, and genetic screening are included. 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 invaluable, CRISPR Gene Editing: Methods and Protocols will assist undergraduates, graduates, and researchers with detailed guidelines and methods for the vitally important CRISPR gene editing field. Chapter 3 is available open access under a CC BY 4.0 license via link.springer.com.
Download or read book Pichia Protocols written by James M Cregg and published by Springer Science & Business Media. This book was released on 2007-08-08 with total page 553 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by the methods of restriction enzyme-mediated integration (REMI). Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system. This volume fully updates and expands upon the first edition.
Download or read book Transgenic Mouse Methods and Protocols written by Marten H. Hofker and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 382 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book Transgenesis Techniques written by Alan R. Clarke and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 346 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book Innate Lymphoid Cells written by Shoba Amarnath and published by Humana. This book was released on 2020-03-09 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume explores the latest techniques used to characterize murine and human Innate Lymphoid Cells (ILC) subsets. This book also aims to discuss useful phenotypical and functional assays that may be relevant for studying ILC function in mice and humans. Chapters cover topics such as methods to study microbiome and enteroid cultures; characterizing ILC precursors; and cellular engineering of ILCs for therapeutic use. 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. Cutting-edge and practical, Innate Lymphoid Cells: Methods and Protocols is a valuable research tool for anyone interested in ILC biology, and those interested in fundamental immunology, cellular engineering, cell signaling, and clinician scientists.
Download or read book CRISPR Cas Methods written by M. Tofazzal Islam and published by Humana. This book was released on 2021-07-31 with total page 397 pages. Available in PDF, EPUB and Kindle. Book excerpt: This second volume provides new and updated methods detailing advancements in CRISPR-Cas technical protocols. Chapters guide readers through protocols on prime editing, base editing, multiplex editing, editing in cell-free extract, in silico analysis of gRNA secondary structure and CRISPR-diagnosis. Authoritative and cutting-edge, CRISPR-Cas Methods, Volume 2 aims to serves as a laboratory manual providing scientists with a holistic view of CRISPR-Cas methodologies and its practical application for the editing of crop plants, cell lines, nematode and microorganism. The chapter “CRISPR/Cas9-mediated gene editing in human induced pluripotent stem cells” is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
Download or read book Leishmania written by Joachim Clos and published by Humana Press. This book was released on 2019-05-27 with total page 370 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume explores the latest techniques used to study Leishmania and other trypanosomatidae. Chapters in this book cover a wide range of topics such as cultivation of axenic amastigote forms; phylogeny and comparative genomics; and in vitro and in vivo infection models that look at hosts ranging from cell lines to mammalian and arthropode hosts. 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. Unique and cutting-edge, Leishmania: Methods and Protocols is a valuable guide for both established and novice researchers in the Leishmania field. This book will also be useful to researchers working with other protozoa of the Order Trypanosomatide.
Download or read book Microinjection written by Chengyu Liu and published by Humana. This book was released on 2019-12-10 with total page 540 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book Guide to Research Techniques in Neuroscience written by Matt Carter and published by Academic Press. This book was released on 2022-03-26 with total page 416 pages. Available in PDF, EPUB and Kindle. Book excerpt: Modern neuroscience research is inherently multidisciplinary, with a wide variety of cutting edge new techniques to explore multiple levels of investigation. This Third Edition of Guide to Research Techniques in Neuroscience provides a comprehensive overview of classical and cutting edge methods including their utility, limitations, and how data are presented in the literature. This book can be used as an introduction to neuroscience techniques for anyone new to the field or as a reference for any neuroscientist while reading papers or attending talks. Nearly 200 updated full-color illustrations to clearly convey the theory and practice of neuroscience methods Expands on techniques from previous editions and covers many new techniques including in vivo calcium imaging, fiber photometry, RNA-Seq, brain spheroids, CRISPR-Cas9 genome editing, and more Clear, straightforward explanations of each technique for anyone new to the field A broad scope of methods, from noninvasive brain imaging in human subjects, to electrophysiology in animal models, to recombinant DNA technology in test tubes, to transfection of neurons in cell culture Detailed recommendations on where to find protocols and other resources for specific techniques "Walk-through" boxes that guide readers through experiments step-by-step
Download or read book Mouse Genetics written by Shree Ram Singh and published by Humana. This book was released on 2016-09-17 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Mouse Genetics: Methods and Protocols provide selected mouse genetic techniques and their application in modeling varieties of human diseases. The chapters are mainly focused on the generation of different transgenic mice to accomplish the manipulation of genes of interest, tracing cell lineages and modeling human diseases. Composed in the highly successful Methods in Molecular Biology series format, each chapter contains a brief introduction, a list of necessary materials, systematic, readily reproducible methods and a notes section, which shares tips on troubleshooting in order to avoid known pitfalls. Comprehensive and authoritative, Mouse Genetics: Methods and Protocols promises to deliver fundamental techniques and protocols to geneticists, molecular biologists, cell and developmental biologists, students and postdoctoral fellows working in the various disciplines of mouse biology and modeling human disease.
Download or read book Plant and Food Carotenoids written by Manuel Rodríguez-Concepción and published by Humana. This book was released on 2019-11-20 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume provides a comprehensive compilation of techniques and protocols used in plant and food carotenoid research. Chapters guide readers through seven major areas on core enzyme activities, analysis of carotenoid profiles, new imaging tools, synthesis and degradation dynamics, biotechnology, nutrition, and health. 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. Authoritative and cutting-edge, Plant and Food Carotenoids: Methods and Protocols aims to be helpful to researchers of other disciplines that are impacted by carotenoids, including photosynthesis, biotechnology, food science, and nutrition.