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  • 1
  • 2
    Book
    Book
    Cold Spring Harbor, New York :Cold Spring Harbor Laboratory Press,
    UID:
    almahu_BV043273715
    Format: x, 181 Seiten : , Illustrationen, Diagramme.
    ISBN: 978-1-62182-131-1 , 978-1-62182-130-4
    Language: English
    Subjects: Biology
    RVK:
    Keywords: Repetitive DNS ; Gentechnologie ; CRISPR/Cas-Methode ; Aufsatzsammlung ; Laboratory Manuals
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  • 3
    Online Resource
    Online Resource
    Amsterdam [u.a.] : Elsevier, Acad. Press
    UID:
    b3kat_BV042244885
    Format: 1 Online-Ressource , graph. Darst., Tab.
    ISBN: 9780128011850 , 9780128013342
    Series Statement: Methods in enzymology 546
    Language: English
    Subjects: Biology
    RVK:
    Keywords: Genanalyse ; Aufsatzsammlung ; Electronic books ; Aufsatzsammlung
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  • 4
    UID:
    almahu_BV044545502
    Format: xx, 281 Seiten : , Illustrationen.
    ISBN: 978-0-544-71694-0
    Note: Includes bibliographical references and index
    Additional Edition: Erscheint auch als Online-Ausgabe Doudna, Jennifer A., author Crack in creation Boston : Houghton Mifflin Harcourt, 2017 ISBN 9780544716964
    Language: English
    Subjects: Philosophy
    RVK:
    Keywords: Gentechnologie ; Genetischer Code ; CRISPR/Cas-Methode
    Author information: Sternberg, Samuel H.
    Author information: Doudna, Jennifer A., 1964-,
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  • 5
    UID:
    kobvindex_ZLB34070044
    ISBN: 9780544716964
    Content: " A trailblazing biologist grapples with her role in the biggest scientific discovery of our era: a cheap, easy way of rewriting genetic code, with nearly limitless promise and peril . Not since the atomic bomb has a technology so alarmed its inventors that they warned the world about its use. Not, that is, until the spring of 2015, when biologist Jennifer Doudna called for a worldwide moratorium on the use of the new gene-editing tool CRISPR—, revolutionary new technology that she helped create—,o make heritable changes in human embryos. The cheapest, simplest, most effective way of manipulating DNA ever known, CRISPR may well give us the cure to HIV, genetic diseases, and some cancers, and will help address the world's hunger crisis. Yet even the tiniest changes to DNA could have myriad, unforeseeable consequences—,o say nothing of the ethical and societal repercussions of intentionally mutating embryos to create better humans.Writing with fellow researcher Samuel Sternberg, Doudna shares the thrilling story of her discovery, and passionately argues that enormous responsibility comes with the ability to rewrite the code of life. With CRISPR, she shows, we have effectively taken control of evolution. What will we do with this unfathomable power? "
    Content: Rezension(1): "〈a href=http://www.publishersweekly.com target=blank〉〈img src=https://images.contentreserve.com/pw_logo.png alt=Publisher's Weekly border=0 /〉〈/a〉: April 10, 2017 Doudna, professor of biology at UC-Berkeley, and Sternberg, her former graduate student and current collaborator, explain the basics of the potentially revolutionary CRISPR technology, the events leading up to Doudna’s discovery of that technology, and the ethical dilemmas posed by the newfound ability to alter any living being’s genetic composition. The authors describe the biological mechanisms in a way that nonspecialists can appreciate, though the simplistic diagrams scattered throughout add little to the text. They also enthusiastically survey many of the uses to which CRISPR technology has already been applied, noting the great interest by venture capitalists who have already invested well over $1 billion in this technology. Doudna and Sternberg make a clear distinction between manipulating reproductive and non-reproductive cells, since the former can cause permanent evolutionary shifts. The second half of the book delves into the ethical implications arising from this difference, thoughtfully covering effects on both human and non-human species. Though the authors note that science involves both “competition and collaboration,” they avoid discussion of the myriad conflicts that exist in this exciting new field—an absence that makes the rosy picture presented in this otherwise excellent book just a bit too unbelievable. Illus. Agent: Max Brockman, Brockman Inc. " Rezension(2): "〈a href=http://www.kirkusreviews.com target=blank〉〈img src=https://images.contentreserve.com/kirkus_logo.png alt=Kirkus border=0 /〉〈/a〉: Starred review from April 1, 2017 A pair of biochemists offer a fresh examination of the newest and arguably most effective genetic-engineering tool.Biological spectaculars--e.g., genetic engineering, cloned sheep, in vitro fertilization--have produced headlines and bestsellers but flopped where it counts: they don't save many lives. CRISPR (clustered regularly interspaced short palindromic repeats) is changing that, write Doudna (Chemistry and Molecular Biology/Univ. of California,co-author: Molecular Biology: Principles and Practice, 2011, etc.) and Sternberg in this enthusiastic and definitely not dumbed-down account of gene manipulation that, unlike earlier methods, is precise and easy. In the first half of the book, The Tool, the authors summarize a century of research but focus on the discovery, in the early 2000s, that bacteria possess an ingenious immune system that destroys invading viruses by cutting their DNA into pieces. Within the past decade, researchers converted this into an ingenious technique for literally debugging DNA: putting in good genes in the place of bad. Because CRISPR allows precise and relatively straightforward DNA editing, write the authors, it has transformed every genetic disease--at least, every disease for which we know the underlying mutation--into a potentially treatable target. The second half, The Task, describes the miraculous powers of CRISPR to cure disease and control evolution--but not yet. Replacing a single defective gene cures muscular dystrophy in mice,clinical trials in humans for this and similar disorders (sickle-cell, hemophilia, cystic fibrosis) are in the works. CRISPR can't yet cure cancer, prevent AIDS, wipe out malaria, revive the wooly mammoth, or regenerate a limb, but an avalanche of startups (Doudna's included) is betting billions that it eventually will. An important book about a major scientific advance but not for the faint of heart. Readers not up to speed on high school biology should prepare themselves with a good popular primer on DNA, such as Matthew Cobb's Life's Greatest Secret (2015). COPYRIGHT(2017) Kirkus Reviews, ALL RIGHTS RESERVED. "
    Language: English
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  • 6
    UID:
    almahu_BV044486569
    Format: xvii, 281 Seiten : , Illustrationen.
    ISBN: 978-1-84792-382-0 , 978-1-84792-381-3
    Language: English
    Subjects: Biology
    RVK:
    RVK:
    Keywords: Gentechnologie ; Genetischer Code
    Author information: Sternberg, Samuel H.
    Author information: Doudna, Jennifer A., 1964-,
    Library Location Call Number Volume/Issue/Year Availability
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  • 7
    UID:
    almahu_BV042541968
    Format: getr. Zählung : , Ill., graph. Darst.
    Edition: 2. ed., 1. print.
    ISBN: 978-1-4641-2614-7
    Language: English
    Subjects: Biology
    RVK:
    Keywords: Molekularbiologie
    Author information: Cox, Michael M.
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  • 8
    UID:
    edoccha_9958128010602883
    Format: 1 online resource (594 p.)
    Edition: First edition.
    ISBN: 0-12-801334-6
    Series Statement: Methods in Enzymology, Volume 546
    Content: This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers recent research and methods development for changing the DNA sequence within the genomes of cells and organisms. Focusing on enzymes that generate double-strand breaks in DNA, the chapters describe use of molecular tools to introduce or delete genetic information at specific sites in the genomes of animal, plant and bacterial cells.Continues the legacy of this premier serial with quality chapters authored by leaders in the fieldCovers r
    Note: Description based upon print version of record. , Front Cover; The Use of CRISPR/Cas9, ZFNs, and TALENs in Generating Site-Specific Genome Alterations; Copyright; Contents; Contributors; Preface; Chapter One: In Vitro Enzymology of Cas9; 1. Introduction; 2. Expression and Purification of Cas9; Day 1: Cell transformation; Day 2: Culture growth and induction; Day 3: Cas9 purification by IMAC; Day 4: IEX and SEC chromatographic steps; Day 5: Concentration and storage; 3. Preparation of Guide RNAs; Day 1: Preparation of transcription template; Day 2: In vitro transcription and gel purification; Day 3: Gel purification-continued , 4. Endonuclease Cleavage AssaysSubstrate preparation; Cleavage assay; Interpretation of cleavage assays; 5. Concluding Remarks; Acknowledgments; References; Chapter Two: Targeted Genome Editing in Human Cells Using CRISPR/Cas Nucleases and Truncated Guide RNAs; 1. Introduction; 2. Methods; 2.1. Identification of target sites using ZiFiT; Required materials; Ensure query sequence is valid; Design target sites; 2.2. Construction of tru-gRNA expression plasmids; 2.2.1. Reagents; 2.2.2. Protocol; 2.3. Transfection of sgRNA and Cas9 expression plasmids into human cells; 2.3.1. Reagents , 2.3.2. Protocol2.3.2.1. Prior to Day 1; 2.4. Quantitative T7EI assays to assess frequencies of targeted genome editing; 2.4.1. Reagents; 2.4.2. Protocol; Conflict of Interest; References; Chapter Three: Determining the Specificities of TALENs, Cas9, and Other Genome-Editing Enzymes; 1. Introduction; 1.1. Introduction to programmable nucleases for genome editing; 1.2. Overview of methods to study specificity of genome-editing agents; 1.2.1. Discrete off-target site testing; 1.2.2. Genome-wide selections; 1.2.3. Minimally biased selections in vitro and in cells , 1.3. Insights and improvements from ZFN specificity studies1.4. Insights and improvements from TALEN specificity studies; 1.5. Insights and improvements from Cas9 specificity studies; 2. Methods; 2.1. Overview of in vitro selection-based nuclease specificity profiling; 2.2. Pre-selection library design; 2.3. In vitro selection protocol; 2.3.1. Before Day 1: Design and synthesize pre-selection library oligonucleotides; 2.3.2. Day 1: Circularize library oligonucleotides; 2.3.3. Day 2: Confirm circularization of library oligonucleotides and perform rolling-circle amplification , 2.3.4. Day 3: Quantify and digest pre-selection library2.3.5. Day 4: PCR of pre- and post-selection libraries; 2.3.6. Day 5: High-throughput sequencing and analysis; 2.4. Confirmation of in vitro-identified genomic off-target sites; 3. Conclusion; Acknowledgments; References; Chapter Four: Genome Engineering with Custom Recombinases; 1. Introduction; 2. Target Identification; 3. Recombinase Construction; 4. Measurements of Recombinase Activity; 4.1. Reporter plasmid construction; 4.2. Luciferase assay; 5. Site-Specific Integration; 5.1. Donor plasmid construction; 5.2. Cell culture methods , 5.2.1. PCR confirmation of integration , English
    Additional Edition: ISBN 0-12-801185-8
    Language: English
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  • 9
    UID:
    almahu_9949697543902882
    Format: 1 online resource (594 p.)
    Edition: First edition.
    ISBN: 0-12-801334-6
    Series Statement: Methods in Enzymology, Volume 546
    Content: This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers recent research and methods development for changing the DNA sequence within the genomes of cells and organisms. Focusing on enzymes that generate double-strand breaks in DNA, the chapters describe use of molecular tools to introduce or delete genetic information at specific sites in the genomes of animal, plant and bacterial cells.Continues the legacy of this premier serial with quality chapters authored by leaders in the fieldCovers r
    Note: Description based upon print version of record. , Front Cover; The Use of CRISPR/Cas9, ZFNs, and TALENs in Generating Site-Specific Genome Alterations; Copyright; Contents; Contributors; Preface; Chapter One: In Vitro Enzymology of Cas9; 1. Introduction; 2. Expression and Purification of Cas9; Day 1: Cell transformation; Day 2: Culture growth and induction; Day 3: Cas9 purification by IMAC; Day 4: IEX and SEC chromatographic steps; Day 5: Concentration and storage; 3. Preparation of Guide RNAs; Day 1: Preparation of transcription template; Day 2: In vitro transcription and gel purification; Day 3: Gel purification-continued , 4. Endonuclease Cleavage AssaysSubstrate preparation; Cleavage assay; Interpretation of cleavage assays; 5. Concluding Remarks; Acknowledgments; References; Chapter Two: Targeted Genome Editing in Human Cells Using CRISPR/Cas Nucleases and Truncated Guide RNAs; 1. Introduction; 2. Methods; 2.1. Identification of target sites using ZiFiT; Required materials; Ensure query sequence is valid; Design target sites; 2.2. Construction of tru-gRNA expression plasmids; 2.2.1. Reagents; 2.2.2. Protocol; 2.3. Transfection of sgRNA and Cas9 expression plasmids into human cells; 2.3.1. Reagents , 2.3.2. Protocol2.3.2.1. Prior to Day 1; 2.4. Quantitative T7EI assays to assess frequencies of targeted genome editing; 2.4.1. Reagents; 2.4.2. Protocol; Conflict of Interest; References; Chapter Three: Determining the Specificities of TALENs, Cas9, and Other Genome-Editing Enzymes; 1. Introduction; 1.1. Introduction to programmable nucleases for genome editing; 1.2. Overview of methods to study specificity of genome-editing agents; 1.2.1. Discrete off-target site testing; 1.2.2. Genome-wide selections; 1.2.3. Minimally biased selections in vitro and in cells , 1.3. Insights and improvements from ZFN specificity studies1.4. Insights and improvements from TALEN specificity studies; 1.5. Insights and improvements from Cas9 specificity studies; 2. Methods; 2.1. Overview of in vitro selection-based nuclease specificity profiling; 2.2. Pre-selection library design; 2.3. In vitro selection protocol; 2.3.1. Before Day 1: Design and synthesize pre-selection library oligonucleotides; 2.3.2. Day 1: Circularize library oligonucleotides; 2.3.3. Day 2: Confirm circularization of library oligonucleotides and perform rolling-circle amplification , 2.3.4. Day 3: Quantify and digest pre-selection library2.3.5. Day 4: PCR of pre- and post-selection libraries; 2.3.6. Day 5: High-throughput sequencing and analysis; 2.4. Confirmation of in vitro-identified genomic off-target sites; 3. Conclusion; Acknowledgments; References; Chapter Four: Genome Engineering with Custom Recombinases; 1. Introduction; 2. Target Identification; 3. Recombinase Construction; 4. Measurements of Recombinase Activity; 4.1. Reporter plasmid construction; 4.2. Luciferase assay; 5. Site-Specific Integration; 5.1. Donor plasmid construction; 5.2. Cell culture methods , 5.2.1. PCR confirmation of integration , English
    Additional Edition: ISBN 0-12-801185-8
    Language: English
    Library Location Call Number Volume/Issue/Year Availability
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  • 10
    UID:
    edoccha_9958130955602883
    Format: 1 online resource (595 p.)
    ISBN: 0-12-801615-9
    Series Statement: Methods in Enzymology ; Volume 546
    Content: This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. Methods to assess mitochondrial function is of great interest to neuroscientists studying chronic forms of neurodegeneration, including Parkinson's, Alzheimer's, ALS, Huntington's and other triplet repeat diseases, but also to those working on acute conditions such as stroke and traumatic brain injury. This volume covers research methods on how to assess the life cycle of mitochondria including trafficking, fusion, fission, and degradation. Multiple persp
    Note: Description based upon print version of record. , Front Cover; The Use of CRISPR/Cas9, ZFNs, and TALENs in Generating Site-Specific Genome Alterations; Copyright; Contents; Contributors; Preface; Chapter One: In Vitro Enzymology of Cas9; 1. Introduction; 2. Expression and Purification of Cas9; Day 1: Cell transformation; Day 2: Culture growth and induction; Day 3: Cas9 purification by IMAC; Day 4: IEX and SEC chromatographic steps; Day 5: Concentration and storage; 3. Preparation of Guide RNAs; Day 1: Preparation of transcription template; Day 2: In vitro transcription and gel purification; Day 3: Gel purification-continued , 4. Endonuclease Cleavage AssaysSubstrate preparation; Cleavage assay; Interpretation of cleavage assays; 5. Concluding Remarks; Acknowledgments; References; Chapter Two: Targeted Genome Editing in Human Cells Using CRISPR/Cas Nucleases and Truncated Guide RNAs; 1. Introduction; 2. Methods; 2.1. Identification of target sites using ZiFiT; Required materials; Ensure query sequence is valid; Design target sites; 2.2. Construction of tru-gRNA expression plasmids; 2.2.1. Reagents; 2.2.2. Protocol; 2.3. Transfection of sgRNA and Cas9 expression plasmids into human cells; 2.3.1. Reagents , 2.3.2. Protocol2.3.2.1. Prior to Day 1; 2.4. Quantitative T7EI assays to assess frequencies of targeted genome editing; 2.4.1. Reagents; 2.4.2. Protocol; Conflict of Interest; References; Chapter Three: Determining the Specificities of TALENs, Cas9, and Other Genome-Editing Enzymes; 1. Introduction; 1.1. Introduction to programmable nucleases for genome editing; 1.2. Overview of methods to study specificity of genome-editing agents; 1.2.1. Discrete off-target site testing; 1.2.2. Genome-wide selections; 1.2.3. Minimally biased selections in vitro and in cells , 1.3. Insights and improvements from ZFN specificity studies1.4. Insights and improvements from TALEN specificity studies; 1.5. Insights and improvements from Cas9 specificity studies; 2. Methods; 2.1. Overview of in vitro selection-based nuclease specificity profiling; 2.2. Pre-selection library design; 2.3. In vitro selection protocol; 2.3.1. Before Day 1: Design and synthesize pre-selection library oligonucleotides; 2.3.2. Day 1: Circularize library oligonucleotides; 2.3.3. Day 2: Confirm circularization of library oligonucleotides and perform rolling-circle amplification , 2.3.4. Day 3: Quantify and digest pre-selection library2.3.5. Day 4: PCR of pre- and post-selection libraries; 2.3.6. Day 5: High-throughput sequencing and analysis; 2.4. Confirmation of in vitro-identified genomic off-target sites; 3. Conclusion; Acknowledgments; References; Chapter Four: Genome Engineering with Custom Recombinases; 1. Introduction; 2. Target Identification; 3. Recombinase Construction; 4. Measurements of Recombinase Activity; 4.1. Reporter plasmid construction; 4.2. Luciferase assay; 5. Site-Specific Integration; 5.1. Donor plasmid construction; 5.2. Cell culture methods , 5.2.1. PCR confirmation of integration , English
    Additional Edition: ISBN 0-12-801415-6
    Language: English
    Library Location Call Number Volume/Issue/Year Availability
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