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This second edition provides new and updated protocols that can be used for generation of knockout animals. Chapters guide the reader through basic protocols for three genome editing technologies, target design tools, and specific protocols for each animal. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.Authoritative and cutting-edge, Genome Editing in Animals: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field.
Construction and Evaluation of Zinc Finger Nucleases
Updated Overview of TALEN Construction Systems
CRISPR/Cas9
SNPD-CRISPR: Single Nucleotide Polymorphism-Distinguishable Repression or Enhancement of a Target Gene Expression by CRISPR System
Utilizing Large Functional and Population Genomics Resources for CRISPR/Cas Perturbation Experiment Design
Generation of Genome-Edited Mice by Cytoplasmic Injection of CRISPR-Cas9 RNA
Gene Targeting in Mouse Embryonic Stem Cells via CRISPR/Cas9 Ribonucleoprotein (RNP)-Mediated Genome Editing
Generation of Knock-In Mouse by Genome Editing
Introduction of Genetic Mutations Into Mice by Base Editor and Target-AID
Genome Editing in Mouse and Rat by Electroporation
Generation of Floxed Mice by Sequential Electroporation
Efficient Detection of Flox Mice Using In Vitro Cre Recombination
VCre/VloxP and SCre/SloxP as Reliable Site-Specific Recombination Systems for Genome Engineering
Efficient CRISPR/Cas9-Assisted Knockin of Large DNA Donors by Pronuclear Microinjection During S-Phase in Mouse Zygotes
Genome Editing of Murine Liver Hepatocytes by AAV Vector-Mediated Expression of Cas9 In Vivo
Nonviral Ex Vivo Genome Editing in Mouse Bona Fide Hematopoietic Stem Cells with CRISPR/Cas9
Genome Editing of Rat
A Simple and Efficient Method for Generating KO Rats Using In Vitro Fertilized Oocytes
Editing the Genome of the Golden Hamster (Mesocricetus auratus)
Gene Targeting in Rabbits: Single-Step Generation of Knockout Rabbits by Microinjection of CRISPR/Cas9 Plasmids
Genome Editing of Pig
GEEP Method: An Optimized Electroporation-Mediated Gene Editing Approach for Establishment of Knockout Pig Lines
Genome Editing Mediated by Primordial Germ Cell in Chicken
CRISPR-Cas9-Mediated Genome Modifications in Zebrafish
Genome Editing of Medaka
CRISPR-Cas9-Based Functional Analysis in Amphibians: Xenopus laevis, Xenopus tropicalis, and Pleurodeles waltl
Genome Editing of Silkworms
Improved Genome Editing in the Ascidian Ciona with CRISPR/Cas9 and TALEN
Genome Editing of C. elegans