Allophenic

Allophenic individuals are composed of cells of two different genotypes (often called mosaics or chimeras). Allophenic mice are the basis of the revolution in mouse genetics, allowing mice with specific gene replacements to be generated and studied. Allophenic mice are made by mixing cells from two embryos of different genotypes (1). These new composite embryos are implanted into foster mothers and allowed to develop. The early embryos are able to compensate and form a single individual from the mixture of embryonic cells. The mosaic progeny that result from these manipulations have tissues that contain cells of the two different genotypes. The allophenic mice have also been called tetraparental mice. When the germ cells are also of mixed origin, progeny can be recovered from both genotypes.

 The ability to make allophenic mice in the laboratory was first used to study the contributions of cells to individual tissues. The number of cells that give rise to a particular tissue can be estimated from the proportions of the two genotypes in a large number of allophenic mice. The cellular autonomy of mutant phenotypes can be determined by generating allophenic mice between mutant and wild-type mouse embryos. The two cell types usually differ both in the mutant of interest and in some marker genotype, such as an enzyme polymorphism.

 Two advances in the technology of allophenic mice have contributed to a revolution in mouse genetics in the last few years. The first advance was the ability to use embryonic teratocarcinoma cells from cell culture as one of the two cell types used to make the allophenic mice (2). The cultured cells are injected into the interior of a normal mouse embryo and are incorporated into the embryo, to form an allophenic mouse. The teratocarcinoma cells are totipotent and can even form germ cells that give rise to the gametes for the next generation. The second major advance was the development of techniques for site-directed mutagenesis and gene replacement in mammalian cells in culture (3) (see Gene Targeting). A gene of interest is altered in a specific way in teratocarcinoma cells in culture. A clone of cells with the altered genotype is produced, and cells from the clone are injected into early mouse embryos to produce allophenic mice. These allophenic mice can be studied as mosaic individuals or can be allowed to develop to fertile adults. If some of the germ cells in the adult allophenic mice are derived from the genetically altered teratocarcinoma cells, progeny can be recovered from the gametes produced by the teratocarcinoma cells and used to found a mutant strain of the specific genotype desired. This has led to the production of hundreds of new mouse strains mutant for developmentally important genes. It has also made the construction of mouse models for human disease far easier than in the past. These technologies have been extended from mice to a variety of other mammals; and they could be used to alter the human genome genetically, with far-reaching consequences.

 References

1. B. Mintz (1962) Science 138, 594. 

2. B. Mintz and K. Illmensee (1975) Proc. Natl. Acad. Sci. USA 72, 3585–3589.

3. M. R. Capecchi (1989) Science 244, 1288–1292.

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مواضيع ذات صلة

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