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Genome Editing in Drug Discovery
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Страница 1
Table of Contents
List of Tables
List of Illustrations
Guide
Pages
Genome Editing in Drug Discovery
Страница 8
Preface
List of Abbreviations
List of Contributors
Part 1 Introduction to Drug Discovery and Genome Editing Methods
1 Genome Editing in Drug Discovery
1.1 Introduction
1.2 Genome Engineering
1.3 CRISPR/Cas9
1.4 Applications of CRISPR Cas9 in Drug Discovery
1.5 Concluding Comments
References
2 Historical Overview of Genome Editing from Bacteria to Higher Eukaryotes
2.1 Introduction
2.2 Bacterial DNA Engineering (Recombineering)
2.3 BAC Recombineering
2.4 Metabolic Engineering
2.5 Genetic Engineering in Higher Eukaryotes
2.6 Targeted Endonucleases
2.7 Novel Genome Editing Technologies
2.8 Conclusions
References
3 CRISPR Cas: From Bacterial Adaptive Immunity to the Swiss Army Knife of Drug Discovery
3.1 Introduction
3.2 CRISPR Biology in a Nutshell
3.3 The Diversity of CRISPR Systems
3.3.1 crRNA Biogenesis
3.3.2 Interference 3.3.2.1 Class 1
3.3.2.1.1 Type I
3.3.2.1.2 Type III
3.3.2.1.3 Type IV
3.3.2.2 Class 2
3.3.2.2.1 Type II
3.3.2.2.2
Type V
3.3.2.2.3 Type VI
3.3.3 Adaptation
3.4 CRISPR Systems as the Basis for New Tools in Drug Discovery
3.4.1 Cas Proteins for Gene Editing
3.4.1.1 Cas9 Variants
3.4.1.2 Cas9 Orthologs
3.4.1.3 The Use of Other Cas Proteins in Genome Editing
3.4.2 Application of Cas Proteins Beyond Genome Editing 3.4.2.1 dCas9 Fusions
3.4.2.2 RNA Targeting
3.4.2.3 Biochemical Detection
3.5 Concluding Remarks
References
4 Commercially Available Reagents and Contract Research Services for CRISPR‐Based Studies
4.1 Introduction
4.2 CRISPR Resources and Reagents for Bespoke Editing and Genetic Screening 4.2.1 Publicly Available Resources
4.2.2 Cas9 Enzymes
4.2.3 Cas9 Alternatives
4.2.4 Guide RNA Formats and Reagents
4.2.5 CRISPR Libraries: CRISPR KO, CRISPRa, CRISPRi
4.3
In vivo
CRISPR Screening 4.3.1 Pooled
In vivo
CRISPR Screening in Rodent Models
4.3.2 Considerations for Practice of
in vivo
CRISPR Screening
4.3.3 Next‐Generation
in vivo
CRISPR Screening
4.4 Working with Service Providers for Outsourcing CRISPR Studies
4.4.1 Critical Steps for Outsourcing
4.5 Considerations on Experimental Design and Controls Required when Outsourcing
4.5.1 Considerations on Selecting the Appropriate Cellular Host
4.5.2 Considerations on the Gene/Locus to be Edited
4.5.3 Controlling CRISPR Off‐Target Effects (OTEs) and Clonal Variations
4.5.4 Deciding on Specific Quality Control Experiments on Engineered Cells
4.5.4.1 Confirmation of Gene KO at Protein Level
4.5.4.2 Confirmation of Genetic Manipulation at RNA Level
4.6 Summary
Acknowledgments
References
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