Gene Knockout Protocols

Author : Martin J. Tymms,Ismail Kola
Publisher : Springer Science & Business Media
Page : 434 pages
File Size : 46,8 Mb
Release : 2008-02-03
Category : Medical
ISBN : 9781592592203

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Gene Knockout Protocols by Martin J. Tymms,Ismail Kola Pdf

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.

Author : Ralf Kühn,Wolfgang Wurst
Publisher : Humana Press
Page : 0 pages
File Size : 43,7 Mb
Release : 2009-03-27
Category : Science
ISBN : 1934115266

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Gene Knockout Protocols by Ralf Kühn,Wolfgang Wurst Pdf

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 : John M. Walker
Publisher : Unknown
Page : 128 pages
File Size : 49,9 Mb
Release : 1984
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
ISBN : LCCN:84015696

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Methods in Molecular Biology: Gene knockout protocols by John M. Walker Pdf

Author : Eric B. Kmiec
Publisher : Unknown
Page : 244 pages
File Size : 48,9 Mb
Release : 2000
Category : Gene targeting
ISBN : 1592592155

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Gene Targeting Protocols by Eric B. Kmiec Pdf

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.

Author : Izuho Hatada
Publisher : Springer Nature
Page : 396 pages
File Size : 43,7 Mb
Release : 2023-02-11
Category : Science
ISBN : 9781071630167

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Genome Editing in Animals by Izuho Hatada Pdf

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.

Author : Marten H. Hofker,Jan van Deursen
Publisher : Springer Science & Business Media
Page : 382 pages
File Size : 44,9 Mb
Release : 2008-02-04
Category : Science
ISBN : 9781592593408

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Transgenic Mouse Methods and Protocols by Marten H. Hofker,Jan van Deursen Pdf

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 : Shoba Amarnath
Publisher : Humana
Page : 0 pages
File Size : 43,9 Mb
Release : 2020-03-09
Category : Medical
ISBN : 107160337X

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Innate Lymphoid Cells by Shoba Amarnath Pdf

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.

Author : Yonglun Luo
Publisher : Unknown
Page : 362 pages
File Size : 46,7 Mb
Release : 2019
Category : CRISPR-associated protein 9
ISBN : 1493991701

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CRISPR Gene Editing by Yonglun Luo Pdf

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.

Author : James M Cregg
Publisher : Springer Science & Business Media
Page : 553 pages
File Size : 40,9 Mb
Release : 2007-08-08
Category : Science
ISBN : 9781588294296

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Pichia Protocols by James M Cregg Pdf

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.

Author : Yiping Qi
Publisher : Springer Nature
Page : 360 pages
File Size : 50,8 Mb
Release : 2019-01-04
Category : Science
ISBN : 9781493989911

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Plant Genome Editing with CRISPR Systems by Yiping Qi Pdf

This volume provides readers with wide-ranging coverage of CRISPR systems and their applications in various plant species. The chapters in this book discuss topics such as plant DNA repair and genome editing; analysis of CRISPR-induced mutations; multiplexed CRISPR/Cas9 systems; CRISPR-Cas12a (Cpf1) editing systems; and non-agrobacterium based CRISPR delivery systems. 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 thorough, Plant Genome Editing with CRISPR Systems: Methods and Protocols is a valuable resource for any researcher interested in learning about and using CRISPR systems in plants.

Author : Rudolf Jaenisch,Feng Zhang,Fred Gage
Publisher : Springer
Page : 0 pages
File Size : 54,6 Mb
Release : 2018-08-11
Category : Medical
ISBN : 3319868012

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Genome Editing in Neurosciences by Rudolf Jaenisch,Feng Zhang,Fred Gage Pdf

This book is open access under a CC BY 4.0 license. CRISPR-Cas9 is a rapid, efficient, versatile and relatively cheap method for dissecting the molecular pathways that are the basis of life, as well as for investigating and potentially rectifying faults in these pathways that result in disease. This book reviews how CRISPR-Cas9 and other genome editing techniques are advancing our understanding of development and function in the nervous system, uncovering the molecular causes of neurological disorders and providing tools for gene therapy.

Author : Chengyu Liu,Yubin Du
Publisher : Unknown
Page : 128 pages
File Size : 41,5 Mb
Release : 2019
Category : Biotechnology
ISBN : 1493988328

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Microinjection by Chengyu Liu,Yubin Du Pdf

Author : Francesca Storici
Publisher : Humana Press
Page : 0 pages
File Size : 43,6 Mb
Release : 2014-02-21
Category : Medical
ISBN : 1627037608

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Gene Correction by Francesca Storici Pdf

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 : M. Tofazzal Islam,Kutubuddin Ali Molla
Publisher : Humana
Page : 397 pages
File Size : 48,6 Mb
Release : 2021-07-31
Category : Science
ISBN : 1071616560

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CRISPR-Cas Methods by M. Tofazzal Islam,Kutubuddin Ali Molla Pdf

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.

Author : William Wu,Helen H. Zhang,Michael J. Welsh,Peter B. Kaufman
Publisher : CRC Press
Page : 576 pages
File Size : 43,8 Mb
Release : 2016-04-19
Category : Medical
ISBN : 9781439848326

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Gene Biotechnology by William Wu,Helen H. Zhang,Michael J. Welsh,Peter B. Kaufman Pdf

Covering state-of-the-art technologies and a broad range of practical applications, the Third Edition of Gene Biotechnology presents tools that researchers and students need to understand and apply today's biotechnology techniques. Many of the currently available books in molecular biology contain only protocol recipes, failing to explain the princ