Adaptable doxycycline-regulated gene expression systems for Drosophila
We have engineered two new versions of the doxycycline (dox) inducible system for use in Drosophila. In the first system, we have used the ubiquitously expressed Drosophila actin5C promoter to express the Tet-Off transactivator (tTA) in all tissue. Induction of a luciferase target transgene begins 6...
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| Veröffentlicht in: | Gene 2001-05, Vol.270 (1), p.103-111 |
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| creator | Stebbins, Michael J Yin, Jerry C.P |
| description | We have engineered two new versions of the doxycycline (dox) inducible system for use in
Drosophila. In the first system, we have used the ubiquitously expressed
Drosophila actin5C promoter to express the Tet-Off transactivator (tTA) in all tissue. Induction of a luciferase target transgene begins 6 h after placing the flies on dox-free food. Feeding drug-free food to mothers results in universal target gene expression in their embryos. Larvae raised on regular food also show robust expression of a target reporter gene. In the second version, we have used the Gal4-UAS system to spatially limit expression of the transactivator. Dox withdrawal results in temporally- and spatially-restricted, inducible expression of luciferase in the adult head and embryo. Both the actin5C and Gal4-UAS versions produce more than 100-fold induction of luciferase in the adult, with virtually no leaky expression in the presence of drug. Reporter gene expression is also undetectable in larvae or embryos from mothers fed dox-containing food. Such tight control may be due to the incorporation of
Drosophila insulator elements (SCS and SCS′) into the transgenic vectors. These systems offer a practical, effective alternative to currently available expression systems in the
Drosophila research community. |
| doi_str_mv | 10.1016/S0378-1119(01)00447-4 |
| format | Article |
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Drosophila. In the first system, we have used the ubiquitously expressed
Drosophila actin5C promoter to express the Tet-Off transactivator (tTA) in all tissue. Induction of a luciferase target transgene begins 6 h after placing the flies on dox-free food. Feeding drug-free food to mothers results in universal target gene expression in their embryos. Larvae raised on regular food also show robust expression of a target reporter gene. In the second version, we have used the Gal4-UAS system to spatially limit expression of the transactivator. Dox withdrawal results in temporally- and spatially-restricted, inducible expression of luciferase in the adult head and embryo. Both the actin5C and Gal4-UAS versions produce more than 100-fold induction of luciferase in the adult, with virtually no leaky expression in the presence of drug. Reporter gene expression is also undetectable in larvae or embryos from mothers fed dox-containing food. Such tight control may be due to the incorporation of
Drosophila insulator elements (SCS and SCS′) into the transgenic vectors. These systems offer a practical, effective alternative to currently available expression systems in the
Drosophila research community.</description><identifier>ISSN: 0378-1119</identifier><identifier>EISSN: 1879-0038</identifier><identifier>DOI: 10.1016/S0378-1119(01)00447-4</identifier><identifier>PMID: 11404007</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>actin5C ; actin5C gene ; Actins - genetics ; Animals ; Animals, Genetically Modified ; Dose-Response Relationship, Drug ; doxycycline ; Doxycycline - pharmacology ; Drosophila ; Drosophila - drug effects ; Drosophila - genetics ; Drosophila - growth & development ; Embryo, Nonmammalian - drug effects ; Embryo, Nonmammalian - metabolism ; Gene Dosage ; Gene Expression Regulation - drug effects ; Gene Expression Regulation, Developmental - drug effects ; Inducible ; Kinetics ; Larva - drug effects ; Larva - metabolism ; Luciferases - drug effects ; Luciferases - genetics ; Luciferases - metabolism ; Promoter Regions, Genetic - genetics ; Recombinant Fusion Proteins - drug effects ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - metabolism ; Spatial/temporal ; Tet-Off transactivator ; Tetracycline ; Trans-Activators - genetics ; Transgenes - genetics ; Transgenic</subject><ispartof>Gene, 2001-05, Vol.270 (1), p.103-111</ispartof><rights>2001 Elsevier Science B.V.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c392t-8d015e7574aa8cc66314a1a817e5bcb33b3c3bd7d767058a7b3fc66ba8517a5b3</citedby><cites>FETCH-LOGICAL-c392t-8d015e7574aa8cc66314a1a817e5bcb33b3c3bd7d767058a7b3fc66ba8517a5b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0378-1119(01)00447-4$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,781,785,3551,27929,27930,46000</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11404007$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Stebbins, Michael J</creatorcontrib><creatorcontrib>Yin, Jerry C.P</creatorcontrib><title>Adaptable doxycycline-regulated gene expression systems for Drosophila</title><title>Gene</title><addtitle>Gene</addtitle><description>We have engineered two new versions of the doxycycline (dox) inducible system for use in
Drosophila. In the first system, we have used the ubiquitously expressed
Drosophila actin5C promoter to express the Tet-Off transactivator (tTA) in all tissue. Induction of a luciferase target transgene begins 6 h after placing the flies on dox-free food. Feeding drug-free food to mothers results in universal target gene expression in their embryos. Larvae raised on regular food also show robust expression of a target reporter gene. In the second version, we have used the Gal4-UAS system to spatially limit expression of the transactivator. Dox withdrawal results in temporally- and spatially-restricted, inducible expression of luciferase in the adult head and embryo. Both the actin5C and Gal4-UAS versions produce more than 100-fold induction of luciferase in the adult, with virtually no leaky expression in the presence of drug. Reporter gene expression is also undetectable in larvae or embryos from mothers fed dox-containing food. Such tight control may be due to the incorporation of
Drosophila insulator elements (SCS and SCS′) into the transgenic vectors. These systems offer a practical, effective alternative to currently available expression systems in the
Drosophila research community.</description><subject>actin5C</subject><subject>actin5C gene</subject><subject>Actins - genetics</subject><subject>Animals</subject><subject>Animals, Genetically Modified</subject><subject>Dose-Response Relationship, Drug</subject><subject>doxycycline</subject><subject>Doxycycline - pharmacology</subject><subject>Drosophila</subject><subject>Drosophila - drug effects</subject><subject>Drosophila - genetics</subject><subject>Drosophila - growth & development</subject><subject>Embryo, Nonmammalian - drug effects</subject><subject>Embryo, Nonmammalian - metabolism</subject><subject>Gene Dosage</subject><subject>Gene Expression Regulation - drug effects</subject><subject>Gene Expression Regulation, Developmental - drug effects</subject><subject>Inducible</subject><subject>Kinetics</subject><subject>Larva - drug effects</subject><subject>Larva - metabolism</subject><subject>Luciferases - drug effects</subject><subject>Luciferases - genetics</subject><subject>Luciferases - metabolism</subject><subject>Promoter Regions, Genetic - genetics</subject><subject>Recombinant Fusion Proteins - drug effects</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Spatial/temporal</subject><subject>Tet-Off transactivator</subject><subject>Tetracycline</subject><subject>Trans-Activators - genetics</subject><subject>Transgenes - genetics</subject><subject>Transgenic</subject><issn>0378-1119</issn><issn>1879-0038</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkMtOwzAQRS0EoqXwCaCsECwCM3VcuytUladUiQWwtvyYFqM0CXaK6N8TaAVLZjObc-dqDmPHCBcIOLp8Ai5VjojjM8BzgKKQebHD-qjkOAfgapf1f5EeO0jpDboRYrjPeogFFACyz24n3jStsSVlvv5cu7UrQ0V5pMWqNC35bEEVZfTZREop1FWW1qmlZcrmdcyuY53q5jWU5pDtzU2Z6Gi7B-zl9uZ5ep_PHu8eppNZ7vh42ObKAwqSQhbGKOdGI46FQaNQkrDOcm6549ZLL0cShDLS8nlHWaMESiMsH7DTzd0m1u8rSq1ehuSoLE1F9SppCeMhl1z9C6JUigsBHSg2oOueSZHmuolhaeJaI-hv0_rHtP7WqAH1j2lddLmTbcHKLsn_pbZqO-BqA1Dn4yNQ1MkFqhz5EMm12tfhn4ovDCSN1g</recordid><startdate>20010530</startdate><enddate>20010530</enddate><creator>Stebbins, Michael J</creator><creator>Yin, Jerry C.P</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7SS</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20010530</creationdate><title>Adaptable doxycycline-regulated gene expression systems for Drosophila</title><author>Stebbins, Michael J ; Yin, Jerry C.P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c392t-8d015e7574aa8cc66314a1a817e5bcb33b3c3bd7d767058a7b3fc66ba8517a5b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>actin5C</topic><topic>actin5C gene</topic><topic>Actins - genetics</topic><topic>Animals</topic><topic>Animals, Genetically Modified</topic><topic>Dose-Response Relationship, Drug</topic><topic>doxycycline</topic><topic>Doxycycline - pharmacology</topic><topic>Drosophila</topic><topic>Drosophila - drug effects</topic><topic>Drosophila - genetics</topic><topic>Drosophila - growth & development</topic><topic>Embryo, Nonmammalian - drug effects</topic><topic>Embryo, Nonmammalian - metabolism</topic><topic>Gene Dosage</topic><topic>Gene Expression Regulation - drug effects</topic><topic>Gene Expression Regulation, Developmental - drug effects</topic><topic>Inducible</topic><topic>Kinetics</topic><topic>Larva - drug effects</topic><topic>Larva - metabolism</topic><topic>Luciferases - drug effects</topic><topic>Luciferases - genetics</topic><topic>Luciferases - metabolism</topic><topic>Promoter Regions, Genetic - genetics</topic><topic>Recombinant Fusion Proteins - drug effects</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Spatial/temporal</topic><topic>Tet-Off transactivator</topic><topic>Tetracycline</topic><topic>Trans-Activators - genetics</topic><topic>Transgenes - genetics</topic><topic>Transgenic</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Stebbins, Michael J</creatorcontrib><creatorcontrib>Yin, Jerry C.P</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Gene</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Stebbins, Michael J</au><au>Yin, Jerry C.P</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Adaptable doxycycline-regulated gene expression systems for Drosophila</atitle><jtitle>Gene</jtitle><addtitle>Gene</addtitle><date>2001-05-30</date><risdate>2001</risdate><volume>270</volume><issue>1</issue><spage>103</spage><epage>111</epage><pages>103-111</pages><issn>0378-1119</issn><eissn>1879-0038</eissn><abstract>We have engineered two new versions of the doxycycline (dox) inducible system for use in
Drosophila. In the first system, we have used the ubiquitously expressed
Drosophila actin5C promoter to express the Tet-Off transactivator (tTA) in all tissue. Induction of a luciferase target transgene begins 6 h after placing the flies on dox-free food. Feeding drug-free food to mothers results in universal target gene expression in their embryos. Larvae raised on regular food also show robust expression of a target reporter gene. In the second version, we have used the Gal4-UAS system to spatially limit expression of the transactivator. Dox withdrawal results in temporally- and spatially-restricted, inducible expression of luciferase in the adult head and embryo. Both the actin5C and Gal4-UAS versions produce more than 100-fold induction of luciferase in the adult, with virtually no leaky expression in the presence of drug. Reporter gene expression is also undetectable in larvae or embryos from mothers fed dox-containing food. Such tight control may be due to the incorporation of
Drosophila insulator elements (SCS and SCS′) into the transgenic vectors. These systems offer a practical, effective alternative to currently available expression systems in the
Drosophila research community.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>11404007</pmid><doi>10.1016/S0378-1119(01)00447-4</doi><tpages>9</tpages></addata></record> |
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| subjects | actin5C actin5C gene Actins - genetics Animals Animals, Genetically Modified Dose-Response Relationship, Drug doxycycline Doxycycline - pharmacology Drosophila Drosophila - drug effects Drosophila - genetics Drosophila - growth & development Embryo, Nonmammalian - drug effects Embryo, Nonmammalian - metabolism Gene Dosage Gene Expression Regulation - drug effects Gene Expression Regulation, Developmental - drug effects Inducible Kinetics Larva - drug effects Larva - metabolism Luciferases - drug effects Luciferases - genetics Luciferases - metabolism Promoter Regions, Genetic - genetics Recombinant Fusion Proteins - drug effects Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Spatial/temporal Tet-Off transactivator Tetracycline Trans-Activators - genetics Transgenes - genetics Transgenic |
| title | Adaptable doxycycline-regulated gene expression systems for Drosophila |
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