Format:
Online-Ressource (XIII, 380 p. 54 illus., 6 illus. in color, digital)
ISBN:
9789400745315
Series Statement:
Topics in Current Genetics 23
Content:
The post-genomic era has brought new challenges and opportunities in all fields of the biology. In this context, several genome engineering technologies have emerged that will help deciphering genes function by as well as improve gene therapy strategies. Genomic modifications such as knock-in, knock-out, knock-down, sequence replacement or modification can today be routinely performed. However, in front of this large palette of methodologies scientists may experience difficulties to gather useful informations scattered within the literature. This book aims to present the state of this field from basic mechanisms of site-directed modifications to their applications in a wide range of organisms such as bacteria, yeast, plants, insects, mammals. It will discuss the problems encountered when using the random integration strategy and present the recent advances made in targeted genome modification. Technologies based on Zinc Finger nucleases, Meganucleases, TALEN, CRE and FLP recombinase, ?C31 integrase, transposases and resolvases are fully detailed with their strengths and weaknesses. All these informations will help students and experienced researchers to understand and choose the best technology for their own purposes.
Note:
Description based upon print version of record
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Site-directed Insertion of Transgenes; Prologue; Contents; Contributors; Part I: Site-Directed Integration of Transgenes; Chapter 1: Transgene Site-Specific Integration: Problems and Solutions; 1.1 Introduction; 1.2 Random Integration and Clonal Dominance: Reality or Myth?; 1.2.1 Viral Vectors and Their Integration Profiles; 1.2.2 Non-viral Vectors and Their Integration Profiles; 1.3 Principal Drawbacks of Gene Transfer Integrative Systems and Solutions; 1.3.1 Vector-on-Host Effects: Insertional Mutagenesis; 1.3.2 Host-on-Vector Effects; 1.3.2.1 Innate Immunity
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1.3.2.2 Transgene Regulation and SilencingChromatin Structure; Epigenetic Modulation; Extrinsic Environmental Factors; 1.3.3 Optimizing the Vector Architecture: Solutions to Ensure Safe and Long-Term Expression; 1.3.3.1 Limiting the Innate Immune Response; 1.3.3.2 Limiting Genotoxicity Using Regulatory Components; 1.3.3.3 Ensure Expression by Modifying Vector Architecture; 1.3.3.4 Number of Insertions Acquired per Cell and Hot Spots of Integration Bias; 1.4 Combining Long-Term Expression and Secure Integration by Specific Locus Targeting: Old Solutions to New Perspectives
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1.4.1 Engineering Integrative Enzymes1.4.1.1 Recombinases; 1.4.1.2 Meganucleases; 1.4.1.3 Zinc-Finger Nucleases; 1.4.1.4 Viral Integrases; 1.4.1.5 Transposases; 1.4.1.6 Other Existing Solutions; 1.4.2 What Defines a Good Insertion Site?; 1.4.2.1 Integration into Repetitive Sequences; Integration into Widely Transcribed Repetitive Genes; Integration into Telomeric Repeats; 1.4.2.2 Integration in Specific Loci; HPRT Locus; Locus ROSA26; AAVsite1; 1.4.2.3 Random Integration; 1.5 Conclusion; References
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Chapter 2: Designing Non-viral Targeted Integrating Vectors for Genome Engineering in Vertebrates2.1 Introduction: Ideal Integrating Vectors and Safe Insertion Sites; 2.2 Zinc Finger Nucleases; 2.3 TALE Nucleases; 2.4 Meganuclease-Based Vectors; 2.5 Site-Specific Recombinase-Based Integrating Vectors; 2.5.1 Site-Specific Tyrosine Recombinases-Based Vectors; 2.5.2 Site-Specific Serine Recombinase, ΦC31-Based Vectors; 2.6 Design of Random Integrating Vectors Based on DNA Transposons and Associated-Targeting Strategy; 2.6.1 Diversity of DNA-Transposon Based Vectors
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2.6.2 Targeting Strategies of DNA Transposon-Based Systems2.6.2.1 Construction of Chimeric Transposases by Fusion to Zinc Finger DBDs; 2.6.2.2 Construction of Bifunctional Targeting Proteins; 2.7 Conclusion; References; Part II: Integration Based on Homologous Recombination; Chapter 3: Gene Targeting and Homologous Recombination in Saccharomyces cerevisiae; 3.1 Introduction; 3.1.1 History; 3.1.2 Outline; 3.2 Transformation with Linearized Plasmids: Targeted Integration; 3.2.1 Ends-in Integration; 3.2.1.1 General Mechanism; 3.2.1.2 Optimizing "Ends-in" Integration
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3.2.1.3 Tagging a Chromosomal Locus in Vivo by "Ends-in" Integration
Additional Edition:
ISBN 9789400745308
Additional Edition:
Buchausg. u.d.T. ISBN 978-94-007-4530-8
Language:
English
Subjects:
Biology
DOI:
10.1007/978-94-007-4531-5
URL:
Volltext
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