Gene Knockout Protocols

Gene Knockout Protocols
Author: Martin J. Tymms,Ismail Kola
Publsiher: Springer Science & Business Media
Total Pages: 431
Release: 2001
Genre: Science
ISBN: 9781592592203

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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
Author: Ralf Kühn,Wolfgang Wurst
Publsiher: Humana Press
Total Pages: 516
Release: 2009-03-27
Genre: Science
ISBN: 1934115266

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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
Author: Martin J. Tymms,Ismail Kola
Publsiher: Unknown
Total Pages: 135
Release: 2001
Genre: Electronic Book
ISBN: OCLC:748540331

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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.

Methods in Molecular Biology Gene knockout protocols

Methods in Molecular Biology  Gene knockout protocols
Author: John M. Walker
Publsiher: Unknown
Total Pages: 135
Release: 1984
Genre: 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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CRISPR Gene Editing

CRISPR Gene Editing
Author: Yonglun Luo
Publsiher: Unknown
Total Pages: 362
Release: 2019
Genre: CRISPR-associated protein 9
ISBN: 1493991701

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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.

Transgenic Mouse

Transgenic Mouse
Author: Marten H. Hofker,Jan van Deursen
Publsiher: Springer Science & Business Media
Total Pages: 374
Release: 2003
Genre: Science
ISBN: 9781592593408

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Genetically very similar to the human species, mice play an important role in biomedical research and have served as experimental models for a wide variety of pathologies, including cancer, cardiovascular diseases, and behavioral disorders. In Transgenic Mouse Methods and Protocols, Marten Hofker and Jan van Deursen have assembled a multidisciplinary collection of readily reproducible methods for working with mice, and particularly for 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. Care is taken to make clear the details of the available approaches, as well as their limitations. Up-to-date and highly practical, Transgenic Mouse Methods and Protocols demonstrates clearly for both novice and expert investigators how to make novel transgenic mouse models, and how to use them effectively to understand the role of gene function in human health and disease.

Leishmania

Leishmania
Author: Joachim Clos
Publsiher: Humana Press
Total Pages: 370
Release: 2019-05-27
Genre: Medical
ISBN: 1493992090

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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 Cas Methods

CRISPR Cas Methods
Author: M. Tofazzal Islam,Kutubuddin Ali Molla
Publsiher: Humana
Total Pages: 397
Release: 2021-07-31
Genre: Science
ISBN: 1071616560

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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.

Pichia Protocols

Pichia Protocols
Author: James M Cregg
Publsiher: Springer Science & Business Media
Total Pages: 268
Release: 2007-08-08
Genre: Science
ISBN: 9781588294296

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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.

CRISPR Cas Systems

CRISPR Cas Systems
Author: Rodolphe Barrangou,John van der Oost
Publsiher: Springer Science & Business Media
Total Pages: 302
Release: 2012-12-13
Genre: Science
ISBN: 9783642346576

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CRISPR/Cas is a recently described defense system that protects bacteria and archaea against invasion by mobile genetic elements such as viruses and plasmids. A wide spectrum of distinct CRISPR/Cas systems has been identified in at least half of the available prokaryotic genomes. On-going structural and functional analyses have resulted in a far greater insight into the functions and possible applications of these systems, although many secrets remain to be discovered. In this book, experts summarize the state of the art in this exciting field.

Microinjection

Microinjection
Author: Chengyu Liu,Yubin Du
Publsiher: Unknown
Total Pages: 540
Release: 2018
Genre: Microinjections
ISBN: 149398831X

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Targeted Genome Editing Using Site Specific Nucleases

Targeted Genome Editing Using Site Specific Nucleases
Author: Takashi Yamamoto
Publsiher: Springer
Total Pages: 205
Release: 2015-01-05
Genre: Science
ISBN: 9784431552277

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This book serves as an introduction to targeted genome editing, beginning with the background of this rapidly developing field and methods for generation of engineered nucleases. Applications of genome editing tools are then described in detail, in iPS cells and diverse organisms such as mice, rats, marine invertebrates, fish, frogs, and plants. Tools that are mentioned include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas9, all of which have received much attention in recent years as breakthrough technologies. Genome editing with engineered nucleases allows us to precisely change the target genome of living cells and is a powerful way to control functional genes. It is feasible in almost all organisms ranging from bacteria to plants and animals, as well as in cultured cells such as ES and iPS cells. Various genome modifications have proven successful, including gene knockout and knock-in experiments with targeting vectors and chromosomal editing. Genome editing technologies hold great promise for the future, for example in biomedical research, clinical medicine, and generation of crops and livestock with desirable traits. A wide range of readers will find this book interesting, and with its focus on applications in a variety of organisms and cells, the book will be valuable for life scientists in all fields.

Gene Targeting Protocols

Gene Targeting Protocols
Author: Eric B. Kmiec
Publsiher: Humana
Total Pages: 233
Release: 2013-08-14
Genre: Medical
ISBN: 1489941584

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The potential now exists in many experimental systems to transfer a cloned, modified gene back into the genome of the host organism. In the ideal situation, the cloned gene is returned to its homologous location in the genome and becomes inserted at the target locus. This process is a controlled means for the repair of DNA damage and ensures accurate chromosome disjunction during meiosis. The paradigm for thinking about the mechanism of this p- cess has emerged primarily from two sources: (1) The principles of reaction mechanics have come from detailed biochemical analyses of the RecA protein purified from Escherichia coli; and (2) the principles of information transfer have been derived from genetic studies carried out in bacteriophage and fungi. A compelling picture of the process of homologous pairing and DNA strand exchange has been influential in directing investigators interested in gene t- geting experiments. The ability to find and pair homologous DNA molecules enables ac- rate gene targeting and is the central phenomenon underlying genetic recombi- tion. Biochemically, the overall process can be thought of as a series of steps in a reaction pathway whereby DNA molecules are brought into homologous register, the four-stranded Holliday structure intermediate is formed, hete- duplex DNA is extended, and DNA strands are exchanged. Not much is known about the biochemical pathway leading to homologous recombination in euka- otes.

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology
Author: Anonim
Publsiher: Unknown
Total Pages: 135
Release: 2022
Genre: Molecular biology
ISBN: 047150338X

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Plant Transcription Factors

Plant Transcription Factors
Author: Nobutoshi Yamaguchi
Publsiher: Humana
Total Pages: 396
Release: 2019-08-08
Genre: Science
ISBN: 1493993569

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This detailed book provides general protocols and technologies that plant biologists worldwide often utilize for the purpose of accelerating research progress in the field of plant transcription factors. Beginning with a brief introduction, the volume continues by exploring methods in the preparation of plant materials, detection of expression levels, interaction tests, and chromatin analyses. 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, Plant Transcription Factors: Methods and Protocols aims to answer a wide range of questions related to transcription factors commonly raised by plant biologists.