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Genome Annotation by Mutation

the savings are not proportionate because more mutant mice must be bred to obtain desired mutants, he says.

Capecchi says the new technique could speed a National Institutes of Health effort to mutate all mouse genes in cell culture and create 900 new lines of mutant mice by 2010 an effort NIH says will be extremely useful for the study of human disease.

Simpler, Faster Mutations in the Whole Mouse Genome Existing gene-targeting methods for deleting large segments of DNA are time-consuming and expensive because they require researchers to make multiple manipulations in mouse embryonic stem cells, which means those cells are less likely to yield mice with the desired stretch of DNA disabled, Capecchi says.

To quickly, cheaply mutate genes or none-gene DNA, Wu and Capecchi used short pieces of DNA known as loxP. Pieces of loxP act like signposts saying, Cut the DNA between these two signposts.

In the new method, loxP is inserted in a chromosome on one mouse, and in a different position on the same chromosome in a second mouse, which also has a gene named Cre in its cells. When the mice are bred, an offspring mouse has loxP DNA on two sites on the same chromosome and also has the Cre gene. The protein made by Cre acts like a knife, cutting the DNA wherever loxP is found.

In the new method, loxP is inserted in a chromosome on one mouse, and in a different position on the same chromosome in a second mouse, which also has a gene named Cre in its cells. When the mice are bred, an offspring mouse has loxP DNA on two sites on the same chromosome and also has the Cre gene. The protein made by Cre acts like a knife, cutting the DNA wherever loxP is found.

The offspring mouse, in turn, is bred with a normal mouse. In a remarkably high average of 10 percent of the offspring, the desired large stretch of DNA is either deleted or duplicated so scientists can see what goes wrong in the mutant mouse and
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