title: Facilitating Neuron-Specific Genetic Manipulations inDrosophila melanogasterUsing a Split GAL4 Repressor.
abstract: Efforts to map neural circuits have been galvanized by the development of genetic technologies that permit the manipulation of targeted sets of neurons in the brains of freely behaving animals. The success of these efforts relies on the experimenter's ability to target arbitrarily small subsets of neurons for manipulation, but such specificity of targeting cannot routinely be achieved using existing methods. InDrosophila melanogaster, a widely-used technique for refined cell type-specific manipulation is the Split GAL4 system, which augments the targeting specificity of the binary GAL4-UAS (Upstream Activating Sequence) system by making GAL4 transcriptional activity contingent upon two enhancers, rather than one. To permit more refined targeting, we introduce here the "Killer Zipper" (KZip+), a suppressor that makes Split GAL4 targeting contingent upon a third enhancer. KZip+acts by disrupting both the formation and activity of Split GAL4 heterodimers, and we show how this added layer of control can be used to selectively remove unwanted cells from a Split GAL4 expression pattern or to subtract neurons of interest from a pattern to determine their requirement in generating a given phenotype. To facilitate application of the KZip+technology, we have developed a versatile set of LexAop-KZip+fly lines that can be used directly with the large number of LexA driver lines with known expression patterns. KZip+significantly sharpens the precision of neuronal genetic control available inDrosophilaand may be extended to other organisms where Split GAL4-like systems are used.
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge Biomedical Campus, CB2 0QH, United Kingdom.|Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147.
authors.forename: Michael-John
authors.initials: MJ
authors.lastname: Dolan
authors.affiliation: Laboratory of Molecular Biology, National Institute of Mental Health, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Haojiang
authors.initials: H
authors.lastname: Luan
authors.affiliation: Laboratory of Molecular Biology, National Institute of Mental Health, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: William C
authors.initials: WC
authors.lastname: Shropshire
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge Biomedical Campus, CB2 0QH, United Kingdom.
authors.forename: Ben
authors.initials: B
authors.lastname: Sutcliffe
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Cocanougher
authors.affiliation: Laboratory of Molecular Biology, National Institute of Mental Health, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Scott
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge Biomedical Campus, CB2 0QH, United Kingdom.
authors.forename: Shahar
authors.initials: S
authors.lastname: Frechter
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147.
authors.forename: Marta
authors.initials: M
authors.lastname: Zlatic
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge Biomedical Campus, CB2 0QH, United Kingdom.
authors.forename: Gregory S X E
authors.identifier: 0000-0002-0587-9355
authors.initials: GSXE
authors.lastname: Jefferis
authors.affiliation: Laboratory of Molecular Biology, National Institute of Mental Health, National Institutes of Health, Bethesda, Maryland 20892 benjaminwhite@mail.nih.gov.
authors.forename: Benjamin H
authors.identifier: 0000-0003-0612-8075
authors.initials: BH
authors.lastname: White
mesh_terms: D000818:Animals; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D029721:Drosophila Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics / Q000254:growth & development; D018507:Gene Expression Regulation, Developmental; D009474:Neurons / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics*; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; C557617:GAL4 protein, Drosophila; D012097:Repressor Proteins; D014157:Transcription Factors
keywords: Drosophila; Gal4-UAS; LexA-LexAop; neural circuits; transgene expression