title: Refined LexA transactivators and their use in combination with the Drosophila Gal4 system.
abstract: The use of binary transcriptional systems offers many advantages for experimentally manipulating gene activity, as exemplified by the success of the Gal4/UAS system in Drosophila. To expand the number of applications, a second independent transactivator (TA) is desirable. Here, we present the optimization of an additional system based on LexA and show how it can be applied. We developed a series of LexA TAs, selectively suppressible via Gal80, that exhibit high transcriptional activity and low detrimental effects when expressed in vivo. In combination with Gal4, an appropriately selected LexA TA permits to program cells with a distinct balance and independent outputs of the two TAs. We demonstrate how the two systems can be combined for manipulating communicating cell populations, converting transient tissue-specific expression patterns into heritable, constitutive activities, and defining cell territories by intersecting TA expression domains. Finally, we describe a versatile enhancer trap system that allows swapping TA and generating mosaics composed of Gal4 and LexA TA-expressing cells. The optimized LexA system facilitates precise analyses of complex biological phenomena and signaling pathways in Drosophila.
authors.affiliation: Institute of Molecular Life Sciences, University of Zurich, CH-8057 Zurich, Switzerland.
authors.forename: Ryohei
authors.initials: R
authors.lastname: Yagi
authors.forename: Franz
authors.initials: F
authors.lastname: Mayer
authors.forename: Konrad
authors.initials: K
authors.lastname: Basler
mesh_terms: D000818:Animals; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D017930:Genes, Reporter; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014921:Wings, Animal / Q000196:embryology / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C118082:brk protein, Drosophila; C064196:dpp protein, Drosophila
references.citation: Nat Rev Genet. 2007 Sep;8(9):663-74
references.pmid: 17703237
references.citation: Proc Natl Acad Sci U S A. 2000 May 9;97(10):5351-6
references.pmid: 10805795
references.citation: Nat Neurosci. 2006 May;9(5):703-9
references.pmid: 16582903
references.citation: Mech Dev. 1999 Feb;80(2):153-8
references.pmid: 10072782
references.citation: Development. 2008 Dec;135(24):4003-13
references.pmid: 19029041
references.citation: Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13543-8
references.pmid: 9811836
references.citation: Nature. 1988 Oct 6;335(6190):563-4
references.pmid: 3047590
references.citation: Methods. 1998 Apr;14(4):421-9
references.pmid: 9608512
references.citation: Cell. 2010 Apr 30;141(3):536-48
references.pmid: 20434990
references.citation: Nature. 1989 Nov 9;342(6246):149-54
references.pmid: 2509940
references.citation: Science. 2003 Dec 5;302(5651):1765-8
references.pmid: 14657498
references.citation: Nucleic Acids Res. 1997 Jul 15;25(14):2723-9
references.pmid: 9207017
references.citation: Cell. 1996 May 3;85(3):357-68
references.pmid: 8616891
references.citation: Proc Natl Acad Sci U S A. 2008 Jan 29;105(4):1255-60
references.pmid: 18202183
references.citation: Proc Natl Acad Sci U S A. 2001 Oct 23;98(22):12596-601
references.pmid: 11675495
references.citation: Cell. 1987 Mar 13;48(5):847-53
references.pmid: 3028647
references.citation: Cell. 2000 Dec 8;103(6):981-91
references.pmid: 11136982
references.citation: Neuron. 1999 Mar;22(3):451-61
references.pmid: 10197526
references.citation: Genetics. 2006 Jun;173(2):769-77
references.pmid: 16547094
references.citation: Genes Dev. 1990 Nov;4(11):2011-23
references.pmid: 2177439
references.citation: Insect Biochem Mol Biol. 2007 Mar;37(3):246-54
references.pmid: 17296499
references.citation: Development. 1997 Jan;124(2):271-8
references.pmid: 9053304
references.citation: EMBO Rep. 2004 Jul;5(7):734-9
references.pmid: 15218525
references.citation: Development. 1993 Jun;118(2):401-15
references.pmid: 8223268
references.citation: Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3312-7
references.pmid: 17360644
references.citation: Genesis. 2002 Sep-Oct;34(1-2):1-15
references.pmid: 12324939
references.citation: Int J Dev Biol. 2002 Jan;46(1):173-6
references.pmid: 11902680