Gene Knockout Protocols Download Ebook PDF Epub Online

Author : Ralf Kühn
Wolfgang Wurst
Publisher : Humana Press
Release : 2014-10-30
Page : 516
Category : Science
ISBN 13 : 9781627038348
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.


Author : Martin J. Tymms
Ismail Kola
Publisher : Springer Science & Business Media
Release : 2001
Page : 431
Category : Science
ISBN 13 : 1592592201
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.


Author : Martin J. Tymms
Ismail Kola
Publisher :
Release : 2001
Page :
Category :
ISBN 13 :
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.


Author : Eric B. Kmiec
Publisher : Humana
Release : 2013-08-14
Page : 233
Category : Medical
ISBN 13 : 9781489941589
Description :


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.


Author : Yonglun Luo
Publisher :
Release : 2019
Page : 362
Category : CRISPR-associated protein 9
ISBN 13 : 9781493991709
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.


Author : Marten H. Hofker
Jan van Deursen
Publisher : Springer Science & Business Media
Release : 2003
Page : 374
Category : Science
ISBN 13 : 1592593402
Description :


A collection of readily reproducible methods for working with mice, and particularly for generating mouse models that will enable you 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. Use effective methods for conditional and inducible gene expression Review mouse models for a wide range of genetic aberrations associated with human disease.


Author : Ralf Kühn
Wolfgang Wurst
Publisher : Humana Press
Release : 2009-03-27
Page : 516
Category : Science
ISBN 13 : 9781934115268
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.


Author : Joachim Clos
Publisher : Humana Press
Release : 2019-05-27
Page : 370
Category : Medical
ISBN 13 : 9781493992096
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.


Author : Sara E. Melville
Publisher : Springer Science & Business Media
Release : 2004
Page : 452
Category : Science
ISBN 13 : 1592597939
Description :


Pioneering bench researchers describe in detail the cutting-edge techniques they have developed for analyzing the genomes and gene products in a diverse range of protozoan and metazoan parasites. These readily reproducible techniques can be used in genomic, functional genomic, and post-genomic studies and include transfection methods and vectors for several protozoan parasites, global analysis using microarrays, gene ablation using RNA interference, gene knockout, mutagenesis, and chromosome manipulation. Some of the protocols require DNA sequence data, whereas others were developed independently of whole-genome sequence data. The protocols presented follow the successful Methods in Molecular BiologyTM series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and the avoidance of known pitfalls.


Author : Damir Janigro
Publisher : Springer Science & Business Media
Release : 2008-01-23
Page : 563
Category : Medical
ISBN 13 : 9781597450218
Description :


Cell Cycle in the Central Nervous System overviews the changes in cell cycle as they relate to prenatal and post natal brain development, progression to neurological disease or tumor formation.Topics covered range from the cell cycle during the prenatal development of the mammalian central nervous system to future directions in postnatal neurogenesis through gene transfer, electrical stimulation, and stem cell introduction. Additional chapters examine the postnatal development of neurons and glia, the regulation of cell cycle in glia, and how that regulation may fail in pretumor conditions or following a nonneoplastic CNS response to injury. Highlights include treatments of the effects of deep brain stimulation on brain development and repair; the connection between the electrophysiological properties of neuroglia, cell cycle, and tumor progression; and the varied immunological responses and their regulation by cell cycle.


Author : Christoph Wittmann
Sang Yup Lee
Publisher : Springer Science & Business Media
Release : 2012-06-15
Page : 388
Category : Medical
ISBN 13 : 9400745346
Description :


Systems Metabolic Engineering is changing the way microbial cell factories are designed and optimized for industrial production. Integrating systems biology and biotechnology with new concepts from synthetic biology enables the global analysis and engineering of microorganisms and bioprocesses at super efficiency and versatility otherwise not accessible. Without doubt, systems metabolic engineering is a major driver towards bio-based production of chemicals, materials and fuels from renewables and thus one of the core technologies of global green growth. In this book, Christoph Wittmann and Sang-Yup Lee have assembled the world leaders on systems metabolic engineering and cover the full story – from genomes and networks via discovery and design to industrial implementation practises. This book is a comprehensive resource for students and researchers from academia and industry interested in systems metabolic engineering. It provides us with the fundaments to targeted engineering of microbial cells for sustainable bio-production and stimulates those who are interested to enter this exiting research field.


Author : M. Tofazzal Islam
Pankaj K. Bhowmik
Publisher : Humana
Release : 2021-07-14
Page : 278
Category : Science
ISBN 13 : 9781071606186
Description :


This volume details the fundamentals of the CRISPR-Cas system, and its protocols illustrate advances in CRISPR-Cas techniques for efficient genome editing. Introductory chapters provide a wide horizon of CRISPR/Cas-based methods and applications. Additional chapters guide readers through HDR-mediated editing, sgRNA design, the step-by-step procedure of multiplex adenine base editing experiments in rice, generating mutants for rice, wheat, Brachypodium, Barley, Flax, and Phytophthora, visual screening of mutants, gene deletion (knock-out), tagging (knock-in) in mammalian cells, the cloning-free (DNA-free) technique, cell-penetrating peptides, generating a genome-edited banana, and nuclear genome editing of Chlamydomonas employing CRISPR-Cpf1 combined with a single-stranded DNA (ssODN) repair template. Authoritative and cutting-edge, CRISPR-Cas Methods aims to assist researchers who are new to the field and are aiming to learn how best to adopt this technology for a particular organism.


Author : Yiping Qi
Publisher : Humana Press
Release : 2019-02-19
Page : 367
Category : Science
ISBN 13 : 9781493989904
Description :


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 : Rocky S. Tuan
Cecilia W. Lo
Publisher : Springer Science & Business Media
Release : 2000-01-21
Page : 522
Category : Science
ISBN 13 : 9780896035768
Description :


This three-volume set, consisting of 142 chapters, is intentionally broad in scope, because of the nature of modern developmental biology.


Author : Z. David Luo
Publisher : Springer Science & Business Media
Release : 2004
Page : 303
Category : Medical
ISBN 13 : 159259770X
Description :


Despite significant advances in molecular biology techniques and in our understanding of the physiology and the behavioral pharmacology of pain transduction, effective, target-specific therapeutic agents for chronic pain are still lacking. In Pain Research: Methods and Protocols, leading researchers who have first-hand experience describe, in step-by-step detail, diverse and novel techniques for dissecting the molecular mechanisms of pain transduction. These readily reproducible methods employ a variety of multidisciplinary approaches ranging from animal pain models and single neutron selection to in vitro single-cell mRNA amplification. The collection includes not only standard, cutting-edge methods, but also novel techniques only recently applied to pain research. The protocols follow the successful Methods in Molecular BiologyTM series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Versatile and easy-to-use, Pain Research: Methods and Protocols offers today's pain researchers in academic and pharmaceutical laboratories powerful tools to unravel the cellular and molecular complexity of pain transduction and set the stage for the next generation of pain medications.


Author : Matt Carter
Jennifer C. Shieh
Publisher : Academic Press
Release : 2015-02-27
Page : 418
Category : Science
ISBN 13 : 0128005971
Description :


Neuroscience is, by definition, a multidisciplinary field: some scientists study genes and proteins at the molecular level while others study neural circuitry using electrophysiology and high-resolution optics. A single topic can be studied using techniques from genetics, imaging, biochemistry, or electrophysiology. Therefore, it can be daunting for young scientists or anyone new to neuroscience to learn how to read the primary literature and develop their own experiments. This volume addresses that gap, gathering multidisciplinary knowledge and providing tools for understanding the neuroscience techniques that are essential to the field, and allowing the reader to design experiments in a variety of neuroscience disciplines. Written to provide a "hands-on" approach for graduate students, postdocs, or anyone new to the neurosciences Techniques within one field are compared, allowing readers to select the best techniques for their own work Includes key articles, books, and protocols for additional detailed study Data analysis boxes in each chapter help with data interpretation and offer guidelines on how best to represent results Walk-through boxes guide readers step-by-step through experiments


Author : Chengyu Liu
Yubin Du
Publisher :
Release : 2018
Page : 540
Category : Microinjections
ISBN 13 : 9781493988310
Description :



Author : Long Jason Lu
Publisher : Humana Press
Release : 2015-01-31
Page : 248
Category : Medical
ISBN 13 : 9781493923977
Description :


This volume opens by covering two main types of approaches widely used to determine essential genes: single-gene knockouts and transposon mutagenesis, in both prokaryotes and Candida albicans. Given the significant advancement in the computational predictions of microbial essential genes, the second half of the book examines four main types of approaches: comparative genomics, supervised machine learning, constraint-based methods, and corrections of transposon mutagenesis data, as well as databases and servers that are often used in studying gene essentiality. Written in the highly successful Methods in Molecular Biology series format, chapters include an introduction 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 up-to-date, Gene Essentiality: Methods and Protocols will aid researchers who wish to further our knowledge in this vital field of study.


Author : Shree Ram Singh
Robert M. Hoffman
Publisher : Humana
Release : 2021-02-20
Page : 217
Category : Science
ISBN 13 : 9781071610077
Description :


This fully updated edition provides 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. 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 up-to-date, Mouse Genetics: Methods and Protocols, Second Edition delivers fundamental techniques and protocols to geneticists, molecular biologists, cell and developmental biologists, students, and postdoctoral fellows working in the various disciplines of genetics, developmental biology, mouse genetics, and modeling human diseases.


Author : Georgios P. D. Argyropoulos
Publisher : Springer Nature
Release : 2020-02-27
Page : 290
Category : Medical
ISBN 13 : 3030356876
Description :


This book provides the first presentation of the state-of-the-art in the application of modern Neuroscience research in predicting, preventing and alleviating the negative sequelae of neurodevelopmental, acquired, or neurodegenerative brain abnormalities on speech and language. To this end, this edited volume brings together contributions from several leading experts in a markedly broad range of disciplines, comprising Neurology, Neurosurgery, Genetics, Engineering, Neuroimaging and Neurostimulation, Neuropsychology, and Speech and Language Therapy.