Gene Transfer and Expression in Animals
Contents Gene engineering technology has been well established in mammalian cells and animals (e.g., mice) as a major revolutionary approach to address a vast spectrum of fundamental biological questions. These include activity assays of promoters, enhancers, and silencers of interest; analysis of expression of mutated genes in their coding regions; analysis of the expression of reporter genes; and applications of gene therapy, using an antisense approach for knocking out the expression of cancer genes or introducing normal genes into patients containing specific mutant genes. Therefore, gene transfer1-7 has a broad range of applications in animal systems. It consists of highly involved techniques that include specific gene isolation and characterization, gene recombination, gene transfer, and analysis of the gene expression in transformed cells or transgenic animals.2,4,8,12 This technology constitutes one of the major advances in medicine and molecular genetics in recent years.1,7,13-15 It has allowed the possibility of treating some human diseases resulting from defects in single genes by transferring normal genes into some of the cells of the patients, a technique known as gene therapy. Some genes that are involved in tumor or cancer formation have been isolated and characterized. This chapter describes detailed procedures for gene transfer and analysis of gene expression in mammalian cells and mice. The techniques can also be adapted for other animal systems.