Methods in Molecular Biology: Gene knockout protocols

Methods in Molecular Biology: Gene knockout protocols PDF Author: John M. Walker
Publisher:
ISBN:
Category : Molecular biology -- v.236. Plant functional genomics -- v.237. G protein signaling -- v.238. Biopolymer methods in tissue engineering -- v.239. Cell migration in inflammation and immunity -- v.240. Mammalian artificial chromosomes -- v.241. Cell cycle checkpoint control protocols -- v.242. Atomic force microscopy -- v.243. Chiral separations: Methods and protocols -- v.244. Protein purification protocols -- v.245-6. Gene delivery to mammalian cells -- v.247. Drosophila cytogenetics protocols -- v.248. Antibody engineering -- v.249. Cytokine protocols -- v.251. HPLC of peptides and proteins: Methods and protocols -- v.265. RNA interference, editing, and modification -- v.274. Photosynthesis research protocols -- v.318. Plant cell culture protocols -- v.323. Arabidopsis protocols
Languages : en
Pages :

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Methods in Molecular Biology: Gene knockout protocols

Methods in Molecular Biology: Gene knockout protocols PDF Author: John M. Walker
Publisher:
ISBN:
Category : Molecular biology -- v.236. Plant functional genomics -- v.237. G protein signaling -- v.238. Biopolymer methods in tissue engineering -- v.239. Cell migration in inflammation and immunity -- v.240. Mammalian artificial chromosomes -- v.241. Cell cycle checkpoint control protocols -- v.242. Atomic force microscopy -- v.243. Chiral separations: Methods and protocols -- v.244. Protein purification protocols -- v.245-6. Gene delivery to mammalian cells -- v.247. Drosophila cytogenetics protocols -- v.248. Antibody engineering -- v.249. Cytokine protocols -- v.251. HPLC of peptides and proteins: Methods and protocols -- v.265. RNA interference, editing, and modification -- v.274. Photosynthesis research protocols -- v.318. Plant cell culture protocols -- v.323. Arabidopsis protocols
Languages : en
Pages :

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Book Description


Gene Knockout Protocols

Gene Knockout Protocols PDF Author: Ralf Kühn
Publisher: Humana Press
ISBN: 9781627038348
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 Knockout Protocols. Methods in Molecular Biology

Gene Knockout Protocols. Methods in Molecular Biology PDF Author: Martin J. Tymms
Publisher:
ISBN:
Category :
Languages : en
Pages :

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Book Description
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.

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.

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.

Mouse Genetics

Mouse Genetics PDF Author: Shree Ram Singh
Publisher: Humana
ISBN: 9781493945672
Category : Medical
Languages : en
Pages : 0

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Book Description
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.

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.

Gene Targeting Protocols

Gene Targeting Protocols PDF Author: Eric B. Kmiec
Publisher:
ISBN: 9780585328560
Category : Science
Languages : en
Pages : 0

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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.

Transgenesis Techniques

Transgenesis Techniques PDF 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.

CRISPR Gene Editing

CRISPR Gene Editing PDF Author: Yonglun Luo
Publisher: Humana Press
ISBN: 9781493991693
Category : Medical
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.