E-Book Overview
Highly skilled investigators with extensive experience in gene targeting and mouse genetics describe their best techniques for the design of targeting constructs and for the analysis of the mouse phenotype. These include embryo transplantation, in vitro embryonic stem cell differentiation, creation of aggregation chimeras, mouse pathology, embryo cryopreservation, and transplantation. State-of-the-art and highly practical, Gene Knockout Protocols not only constitutes an invaluable source of readily reproducible techniques for those just entering the field of gene targeting, but also a key reference for all genetic researchers today.
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Gene Knockouts
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1 Overview Gene Knockouts Paul J. Hertzog and Ismail Kola 1. Introduction The ability to generate a mouse with a targeted mutation in a desired gene has been one of the most important advances in understanding the function of gene products. Not only can gene disruption demonstrate the function of genes, but by disrupting cell development or survival it can also demonstrate the function of specific cell types. Some knockout mice are murine models of human genetic diseases and have proven that a single gene defect is capable of causing disease. In addition, the crossing of mice that are heterozygous or homozygous for specific gene mutations provides important information on human diseases that are caused by multiple genetic defects. In genetic terms, targeted gene disruption is directly complementary to (and arguably more direct than) using mutant strains of mice with a known phenotype to map and identify the diseasecausing genes. The major benefit of gene knockout technology is that it enables the analysis of the function of a protein produced from a specific gene in vivo. Thus, the function can be determined in all normal cell types and the complex interactions between molecules and/or between ce