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Full Description
This detailed book explores how microinjection will be used in the foreseeable future, not only for generating animal models for biomedical research but also for changing economically or ecologically important species that can broadly impact our society in general. The opening half of the book focuses on methods for generating mouse models, as they are still the most popular in genome engineering research, while the second half examines gene-editing in a variety of other species, opened up by the developments in ZFN, TALEN, and CRISPR techniques. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Microinjection: Methods and Protocols serves as an ideal guide for researchers looking to take advantage of the breakthrough technologies in gene-editing and embryo micromanipulations.
Contents
Microinjection and Micromanipulation: A Historical Perspective.- Production of Transgenic Mice by Pronuclear Microinjection.- Transgene Recombineering in Bacterial Artificial Chromosomes.- Using TARGATTTM Technology to Generate Site-Specific Transgenic Mice.- Generating Genetically Engineered Mice Using a Spermatogonial Stem Cell-Mediated Method.- Chimeric Mouse Generation by ES Cell Blastocyst Microinjection and Uterine Transfer.- Creating Knock-In Alleles in Mouse Embryonic Stem Cells by CRISPR/Cas9-Mediated Homologous Recombination without Drug Selection.- Using CRISPR/Cas9 for Gene Knockout in Immunodeficient NSG Mice.- Generation of CRISPR-Edited Rodents Using a Piezo-Driven Zygote Injection Technique.- Delivery of CRISPR-Cas9 into Mouse Zygotes by Electroporation.- Generation of Conditional Knockout Mice by Sequential Insertion of Two loxP Sites In Cis Using CRISPR/Cas9 and Single-Stranded DNA Oligonucleotides.- Improvement of Mouse Cloning from Any Type of Cellby Nuclear Injection.- The CARD Method for Simple Vitrification of Mouse Oocytes: Advantages and Applications.- The CARD Method for Mouse Sperm Cryopreservation and In Vitro Fertilization Using Frozen-Thawed Sperm.- Isolation and Analysis of a Genome-Edited Single-Hepatocyte from a Cas9 Transgenic Mouse Line.- Microinjection and Oviduct Transfer Procedures for Rat Model Generation with CRISPR-Cas9 Technology.- Molecular Aspects of Zinc Finger Nucleases (ZFNs)-Mediated Gene Editing in Rat Embryos.- Organ Generation from Knocked-Out Rat Blastocysts Complemented with Pluripotent Stem Cells.- Generation of Rabbit Models by Gene Editing Nucleases.- Production of Genetically Engineered Porcine Embryos by Handmade Cloning.- Electrofusion of 2-Cell Embryos for Porcine Tetraploid Embryo Production.- Gene Knockouts in Goats Using CRISPR/Cas9 System and Somatic Cell Nuclear Transfer.- Generating Goat Mammary Gland Bioreactors for Producing Recombinant Proteins by Gene Targeting.- Production of Transgenic Chickens Using Cultured Primordial Germ Cells and Gonocytes.- Using Microinjection to Generate Genetically Modified Caenorhabditis elegans by CRISPR/Cas9 Editing.- Microinjection in Zebrafish for Genome Editing and Functional Studies.- Microinjection of Marine Fish Eggs.- Generating Gene Knockout Oryzias Latipes and Rice Field Eel Using TALENs Method.- Functional Studies of Transcriptional Cofactors via Microinjection-Mediated Gene Editing in Xenopus.- Microinjection of Live Mammalian Cells: A Delivery Method that Provides Added Versatility to the Study of Cellular Function.
