Gene Knockout Protocols

Gene Knockout Protocols PDF Author: Martin J. Tymms
Publisher: Springer Science & Business Media
ISBN: 1592592201
Category : Medical
Languages : en
Pages : 434

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Book Description
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.

Gene Knockout Protocols

Gene Knockout Protocols PDF Author: Martin J. Tymms
Publisher: Springer Science & Business Media
ISBN: 1592592201
Category : Medical
Languages : en
Pages : 434

Get Book

Book Description
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.

Gene Knockout Protocols

Gene Knockout Protocols PDF Author: Ralf Kühn
Publisher: Humana Press
ISBN: 9781934115268
Category : Science
Languages : en
Pages : 0

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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.

Gene Targeting Protocols

Gene Targeting Protocols PDF Author: Eric B. Kmiec
Publisher:
ISBN: 9781592592159
Category : Gene targeting
Languages : en
Pages : 244

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Book Description
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.

Pichia Protocols

Pichia Protocols PDF Author: James M Cregg
Publisher: Springer Science & Business Media
ISBN: 1588294293
Category : Science
Languages : en
Pages : 553

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Book Description
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.

Leishmania

Leishmania PDF Author: Joachim Clos
Publisher: Humana Press
ISBN: 9781493992096
Category : Medical
Languages : en
Pages : 370

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Book Description
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.

CRISPR Gene Editing

CRISPR Gene Editing PDF Author: Yonglun Luo
Publisher:
ISBN: 9781493991709
Category : CRISPR-associated protein 9
Languages : en
Pages : 362

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Book Description
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.

Microinjection

Microinjection PDF Author: Chengyu Liu
Publisher: Humana
ISBN: 9781493993987
Category : Science
Languages : en
Pages : 540

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Book Description
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.

Genome Editing in Animals

Genome Editing in Animals PDF Author: Izuho Hatada
Publisher: Springer Nature
ISBN: 1071630164
Category : Science
Languages : en
Pages : 396

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Book Description
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.

Transgenic Mouse Methods and Protocols

Transgenic Mouse Methods and Protocols PDF 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.

Guide to Research Techniques in Neuroscience

Guide to Research Techniques in Neuroscience PDF Author: Matt Carter
Publisher: Academic Press
ISBN: 0323915612
Category : Medical
Languages : en
Pages : 416

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Book Description
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