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Genetics, Vol. 168, 1477-1489, November 2004, Copyright © 2004
doi:10.1534/genetics.104.030882
Gene Deletions by Ends-In Targeting in Drosophila melanogaster
Heng B. Xie and Kent G. Golic1
Department of Biology, University of Utah, Salt Lake City, Utah 84112
1 Corresponding author: Department of Biology, University of Utah, 257 S. 1400 East, Room 201, Salt Lake City, UT 84112.
E-mail: golic{at}biology.utah.edu
Following the advent of a gene targeting technique in Drosophila, different methods have been developed to modify the Drosophila genome. The initial demonstration of gene targeting in flies used an ends-in method, which generates a duplication of the target locus. The duplicated locus can then be efficiently reduced to a single copy by generating a double-strand break between the duplicated segments. This method has been used to knock out target genes by introducing point mutations. A derivative of this method is reported here. By using different homologous regions for the targeting and reduction steps, a complete deletion of the target gene can be generated to produce a definitive null allele. The breakpoints of the deletion can be precisely controlled. Unlike ends-out targeting, this method does not leave exogenous sequence at the deleted locus. Three endogenous genes, Sir2, Sirt2, and p53 have been successfully deleted using this method.
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