E-Book Overview
As the major task of sequencing the human genome nears completion and the full complement of human genes are catalogued, the task of understanding the normal biological functions of genes and how their alteration leads to diseased states becomes more imperative. In Gene Knockout Protocols, highly skilled investigators with extensive experience in gene targeting and mouse genetics describe their best techniques for the design of targeting constructs and for genetic phenotype analysis. These proven methods contain step-by-step instructions, as well as notes on pitfalls to avoid, and emphasize techniques that are relevant to researchers carrying out gene targeting work. These include embryo transplantation, in vitro embryonic stem cell differentiation, creation of aggregation chimeras, mouse pathology, embryo cryopreservation, and transplantation. Issues such as the use of existing mouse mutation resources and the influence of genetic background and epigenetic effects upon phenotype are also covered. 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.
E-Book Content
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