title: CoinFLP: a system for efficient mosaic screening and for visualizing clonal boundaries in Drosophila.
abstract: Screens in mosaic Drosophila tissues that use chemical mutagenesis have identified many regulators of growth and patterning. Many of the mutant phenotypes observed were contingent upon the presence of both wild-type and mutant cells in the same tissue. More recently, large collections of RNAi lines or cDNAs expressed under Gal4/UAS control have been used to alter gene expression uniformly in specific tissues. However, these newer approaches are not easily combined with the efficient generation of genetic mosaics. The CoinFLP system described here enables mosaic screens in the context of gene knockdown or overexpression by automatically generating a reliable ratio of mutant to wild-type tissue in a developmentally controlled manner. CoinFLP-Gal4 generates mosaic tissues composed of clones of which only a subset expresses Gal4. CoinFLP-LexGAD/Gal4 generates tissues composed of clones that express either Gal4 or LexGAD, thus allowing the study of interactions between different types of genetically manipulated cells. By combining CoinFLP-LexGAD/Gal4 with the split-GFP system GRASP, boundaries between genetically distinct cell populations can be visualized at high resolution.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley 361 LSA, Berkeley, CA 94720-3200, USA.
authors.forename: Justin A
authors.initials: JA
authors.lastname: Bosch
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley 361 LSA, Berkeley, CA 94720-3200, USA.
authors.forename: Ngoc Han
authors.initials: NH
authors.lastname: Tran
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley 361 LSA, Berkeley, CA 94720-3200, USA ikh@berkeley.edu.
authors.forename: Iswar K
authors.initials: IK
authors.lastname: Hariharan
mesh_terms: D000818:Animals; D003433:Crosses, Genetic; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins / Q000378:metabolism; D005123:Eye / Q000033:anatomy & histology; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D055879:Gene Knock-In Techniques; D055785:Gene Knockdown Techniques; D057166:High-Throughput Screening Assays / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D060227:Imaginal Discs / Q000033:anatomy & histology; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D009030:Mosaicism; D014157:Transcription Factors / Q000378:metabolism; D014921:Wings, Animal / Q000033:anatomy & histology
chemical_list: D029721:Drosophila Proteins; C557617:GAL4 protein, Drosophila; D014157:Transcription Factors
keywords: Boundary; Drosophila; Mosaic screen; Overexpression; RNAi