title: W::Neo: a novel dual-selection marker for high efficiency gene targeting in Drosophila.
abstract: We have recently developed a so-called genomic engineering approach that allows for directed, efficient and versatile modifications of Drosophila genome by combining the homologous recombination (HR)-based gene targeting with site-specific DNA integration. In genomic engineering and several similar approaches, a "founder" knock-out line must be generated first through HR-based gene targeting, which can still be a potentially time and resource intensive process. To significantly improve the efficiency and success rate of HR-based gene targeting in Drosophila, we have generated a new dual-selection marker termed W::Neo, which is a direct fusion between proteins of eye color marker White (W) and neomycin resistance (Neo). In HR-based gene targeting experiments, mutants carrying W::Neo as the selection marker can be enriched as much as fifty times by taking advantage of the antibiotic selection in Drosophila larvae. We have successfully carried out three independent gene targeting experiments using the W::Neo to generate genomic engineering founder knock-out lines in Drosophila.
authors.affiliation: Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Wenke
authors.initials: W
authors.lastname: Zhou
authors.forename: Juan
authors.initials: J
authors.lastname: Huang
authors.forename: Annie M
authors.initials: AM
authors.lastname: Watson
authors.forename: Yang
authors.initials: Y
authors.lastname: Hong
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004330:Drosophila / Q000235:genetics; D005136:Eye Proteins / Q000235:genetics; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods; D005819:Genetic Markers; D059765:Homologous Recombination
chemical_list: D005136:Eye Proteins; D005819:Genetic Markers