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      Improved and expanded Q-system reagents for genetic manipulations

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          Abstract

          The Q-system is a repressible binary expression system for transgenic manipulations in living organisms. Through protein engineering and in vivo functional tests, we report here new variants of the Q-system transcriptional activator, including QF2, that allows the Q-system to drive strong and ubiquitous expression for the first time in all tissues. Our new QF2, GAL4QF and LexAQF chimeric transcriptional activators substantially enrich the toolkit available for transgenic regulation in Drosophila.

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          The Q system: a repressible binary system for transgene expression, lineage tracing, and mosaic analysis.

          Christopher J. Potter, Bosiljka Tasic, Emilie Russler … (2010)
          We describe a new repressible binary expression system based on the regulatory genes from the Neurospora qa gene cluster. This "Q system" offers attractive features for transgene expression in Drosophila and mammalian cells: low basal expression in the absence of the transcriptional activator QF, high QF-induced expression, and QF repression by its repressor QS. Additionally, feeding flies quinic acid can relieve QS repression. The Q system offers many applications, including (1) intersectional "logic gates" with the GAL4 system for manipulating transgene expression patterns, (2) GAL4-independent MARCM analysis, and (3) coupled MARCM analysis to independently visualize and genetically manipulate siblings from any cell division. We demonstrate the utility of the Q system in determining cell division patterns of a neuronal lineage and gene function in cell growth and proliferation, and in dissecting neurons responsible for olfactory attraction. The Q system can be expanded to other uses in Drosophila and to any organism conducive to transgenesis. 2010 Elsevier Inc. All rights reserved.
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            Genetic mosaic with dual binary transcriptional systems in Drosophila.

            Sen-Lin Lai, Tzumin Lee (2006)
            MARCM (mosaic analysis with a repressible cell marker) involves specific labeling of GAL80-minus and GAL4-positive homozygous cells in otherwise heterozygous tissues. Here we demonstrate how the concurrent use of two independent binary transcriptional systems may facilitate complex MARCM studies in the Drosophila nervous system. By fusing LexA with the VP16 acidic activation domain (VP16) or the GAL4 activation domain (GAD), we obtained both GAL80-insensitive and GAL80-suppressible transcriptional factors. LexA::VP16 can mediate MARCM-independent binary transgene induction in mosaic organisms. The incorporation of LexA::GAD into MARCM, which we call dual-expression-control MARCM, permits the induction of distinct transgenes in different patterns among GAL80-minus cells in mosaic tissues. Lineage analysis with dual-expression-control MARCM suggested the presence of neuroglioblasts in the developing optic lobes but did not indicate the production of glia by postembryonic mushroom body neuronal precursors. In addition, dual-expression-control MARCM with a ubiquitous LexA::GAD driver revealed many unidentified cells in the GAL4-GH146-positive projection neuron lineages.
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              Reiterative use of the EGF receptor triggers differentiation of all cell types in the Drosophila eye.

              M. Freeman (1996)
              The Drosophila eye has contributed much to our knowledge of cell differentiation. This understanding has primarily come from the study of the R7 photoreceptor; much less is known about the development of the other classes of photoreceptor or the nonneuronal cone or pigment cells. I have used a dominant-negative form of the Drosophila epidermal growth factor receptor (DER) to show that this receptor tyrosine kinase (RTK) is required for the differentiation of all these cell types, and I have also shown that DER is sufficient to trigger their development. DER is even required in R7, where it can replace Sevenless, another RTK. These results broaden our view of eye development to include the whole ommatidium and suggest that reiterative activation of DER is critical for triggering the differentiation of all cell types.
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                Author and article information

                Journal
                Journal ID (nlm-journal-id): 101215604
                Journal ID (pubmed-jr-id): 32338
                Journal ID (nlm-ta): Nat Methods
                Journal ID (iso-abbrev): Nat. Methods
                Title: Nature methods
                ISSN (Print): 1548-7091
                ISSN (Electronic): 1548-7105
                Publication date Nihms-submitted: 11 December 2014
                Publication date (Electronic): 12 January 2015
                Publication date (Print): March 2015
                Publication date PMC-release: 01 September 2015
                Volume: 12
                Issue: 3
                Pages: 219-222
                Affiliations
                [1 ]The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA
                [2 ]Howard Hughes Medical Institute, Stanford University, Stanford, CA, USA
                [3 ]Department of Biology, Stanford University, Stanford, CA, USA
                [4 ]Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA
                Author notes
                Correspondence should be addressed to: C.J.P. ( cpotter@ 123456jhmi.edu )
                Article
                Manuscript ID: NIHMS648245
                DOI: 10.1038/nmeth.3250
                PMC ID: 4344399
                PubMed ID: 25581800
                SO-VID: a3092a3a-60b7-452c-a325-719c90066da0
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                ScienceOpen disciplines: Life sciences

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