Gene deletions by ends-in targeting in Drosophila melanogaster.
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.
Xie
Heng B
HB
Department of Biology, University of Utah, Salt Lake City, Utah 84112, USA.
Golic
Kent G
KG
D000818:Animals; D004251:DNA Transposable Elements; D029721:Drosophila Proteins / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics*; D017353:Gene Deletion; D018390:Gene Targeting; D006655:Histone Deacetylases / Q000235:genetics; D037761:Sirtuins / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
D004251:DNA Transposable Elements; D029721:Drosophila Proteins; D016159:Tumor Suppressor Protein p53; C447899:Sirt2 protein, Drosophila; D037761:Sirtuins; D006655:Histone Deacetylases