图书简介
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.- Non-Viral 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.
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