A versatile platform for creating a comprehensive UAS-ORFeome library in Drosophila

• DOI: 10.1242/dev.088757
• Publication Date: 2013-05-02T08:14:21Z
• Authors: Johannes Bischof; Mikael Björklund; Edy Furger; Claus Schertel; Jussi Taipale; Konrad Basler
• Keywords: ORFS; Stop codon; Barcode

Overexpression screens are used to explore gene functions in Drosophila, but this strategy suffers from the lack of comprehensive and systematic fly strain collections efficient methods for generating such collections. Here, we present a that could be efficiently generate large numbers transgenic Drosophila strains, collection 1149 UAS-ORF lines were created with site-specific ΦC31 integrase method. For collection, set 655 genes cloned as two variants, either an open reading frame (ORF) native stop codon or C-terminal 3xHA tag. To streamline procedure generation, demonstrate utility injecting pools plasmids into embryos, each plasmid containing randomised sequence (barcode) serves unique identifier and, subsequently, strains. We also developed swapping technique facilitates rapid exchange promoters epitope tags vivo, expanding versatility ORF collection. The work described here basis library Gal4/UAS-regulated transgenes.