CRISPR in Animals and Animal Models: Volume 152

Cover
Title page
Copyright
Contents
Contributors
Chapter One - CRISPR History: Discovery, Characterization, and Prosperity
	Abstract
	1 Introduction
	2 Discovery of CRISPR-Cas systems
		2.1 Identification of CRISPR Loci and cas Genes
		2.2 Prediction of Small RNA-Based Antiviral Functions of CRISPR
			2.2.1 Identification of CRISPR-Related Small RNAs
			2.2.2 Origin of Spacers in CRISPR Loci
		2.3 Experimental Demonstration of the Basic Mechanism of CRISPR-Cas System
			2.3.1 Adaptation
			2.3.2 crRNA Biogenesis
			2.3.3 Interference
	3 Molecular mechanisms of nucleic acid interference
		3.1 PAM-Dependent DNA Interference by Type I CRISPR-Cas Systems
		3.2 Type II CRISPR-Cas System Requires a Unique tracrRNA for DNA Interference
		3.3 RNA-Activated DNA Interference by Type III CRISPR-Cas Systems
	4 Discovery of anti-CRISPR system
	5 Identification of novel CRISPR-Cas systems
	6 Blooming of CRISPR technology
	References
Chapter Two  - CRISPR/Cas9 Technology: Applications and Human Disease Modeling
	Abstract
	1 Genome engineering tools
		1.1 Genome Engineering Technologies
			1.1.1 Homologous Recombination (HR)
			1.1.2 Double-Strand Breaks (DSBs)
			1.1.3 NHEJ Versus HDR
			1.1.4 Zinc Finger Nuclease (ZFN)
			1.1.5 Transcription Activator-Like Effector Nucleases (TALENs)
			1.1.6 CRISPR/Cas9
		1.2 Off Target Effects
	2 CRISPR applications
		2.1 CRISPRi and CRISPRa: Regulation of Transcriptional Levels
		2.2 Epigenome Edition
		2.3 Genome Imaging
		2.4 Large-Scale Functional Genomic Studies Using CRISPR/Cas9
		2.5 Edition of Single-Stranded RNA
	3 CRISPR applications in biomedical modeling
		3.1 Neurological Disease Models
		3.2 Cancer Models
		3.3 Cardiovascular Disease Models
		3.4 Infectious Disease Models
		3.5 Immunodeficiency Models
	4 Concluding remarks
	References
Chapter Three - Dynamics of Indel Profiles Induced by Various CRISPR/Cas9 Delivery Methods
	Abstract
	1 Introduction
		1.1 Genome Editing and DNA Repair
		1.2 Indel Detection Methodologies
		1.3 IDAA for Indel Detection and gRNA Selection
	2 Materials and methods
		2.1 gRNAs, Plasmid Vectors and Cell Lines
		2.2 Cas9 and gRNA Delivery Conditions
		2.3 Efficiency Measurements
	3 Results
		3.1 Rationale and Design of the Study
		3.2 Transfection and Integration Dynamics
		3.3 Efficiency of Generating Indels Over Time
		3.4 Indel Profiles Over Time
		3.5 Dynamic Effect of gRNA Stability on Indel Efficiency
		3.6 Comparison of IDAA and TIDE Methods
	4 Concluding remarks
	Acknowledgments
	References
Chapter Four - CRISPR Libraries and Screening
	Abstract
	1 Introduction
	2 Mouse models of cancer
	3 CRISPR-Cas9 systems
	4 CRISPR libraries
		4.1 Library Scale and Diversity
		4.2 sgRNA Design
		4.3 Molecular Barcoding
		4.4 Essential Controls
		4.5 DNA Extraction and Library Amplification
	5 Concluding remarks
	References
Chapter Five - Genome Engineering Using Haploid Embryonic Stem Cells
	Abstract
	1 Introduction
	2 Generation of haploid ES cells
	3 Purification and maintenance of haploid cells
	4 Genome editing
		4.1 CRISPR/Cas System
		4.2 Genome Editing in Haploid ES Cells
		4.3 Off-Target Effects
	5 Haploid cells as gamete replacements
	6 Genetic screening using haploid ES cells
	7 Concluding remarks
	References
	Further reading
Chapter Six - CRISPR in Animals and Animal Models
	Abstract
	1 Introduction
		1.1 CRISPR-Cas9 Genome Editing Technology
	2 Methods of producing genetically modified animals
		2.1 Small Model Organisms
		2.2 Rodents
		2.3 Larger Animals
		2.4 Nonhuman Primates
	3 Uses of genetically modified animals
		3.1 Basic Scientific Research
			3.1.1 Rapid Reverse CRISPR Screening
		3.2 Animal Models of Human Diseases
			3.2.1 Cancer
			3.2.2 Genetic Disease and Gene Therapy
		3.3 Industrial Production
			3.3.1 Use of CRISPR in Farming
			3.3.2 Pharmaceutical Production
			3.3.3 Domestic Animals
	4 Looking forward
		4.1 Transgenic Pigs for Xenotransplantation
		4.2 Ecological Engineering
		4.3 Restoring Extinct Species
	5 Ethical considerations
	6 Concluding remarks
	References
Chapter Seven - Gene Editing and CRISPR Therapeutics: Strategies Taught by Cell and Gene Therapy
	Abstract
	1 Genome editors enter the scene
	2 Making advanced technologies ground-breaking
	3 How have we reached this point so quickly?
	4 How far into the clinic will genome-editing go?
	5 A future scenario-risks associated with gene editing businesses
	6 Conclusions
	References
Index
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