Use of Sindbis Virus-Mediated RNA Interference to Demonstrate a Conserved Role of Broad-Complex in Insect Metamorphosis
The transcription factor Broad-Complex (BR-C) is required for differentiation of adult structures as well as for the programmed death of obsolete larval organs during metamorphosis of the fruit fly Drosophila melanogaster. Whether BR-C has a similar role in other holometabolous insects could not be...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2003-12, Vol.100 (26), p.15607-15612 |
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| creator | Uhlirova, Mirka Foy, Brian D. Beaty, Barry J. Olson, Ken E. Riddiford, Lynn M. Jindra, Marek |
| description | The transcription factor Broad-Complex (BR-C) is required for differentiation of adult structures as well as for the programmed death of obsolete larval organs during metamorphosis of the fruit fly Drosophila melanogaster. Whether BR-C has a similar role in other holometabolous insects could not be proven without a loss-of-function genetic test, performed in a non-drosophilid species. Here we use a recombinant Sindbis virus as a tool to silence BR-C expression in the silkmoth Bombyx mori. The virus expressing a BR-C antisense RNA fragment reduced endogenous BR-C mRNA levels in infected tissues (adult wing and leg primordia) via RNA interference (RNAi). The RNAi knock-down of BR-C resulted in the failure of animals to complete the larval-pupal transition or in later morphogenetic defects, including differentiation of adult compound eyes, legs, and wings from their larval progenitors. BR-C RNAi also perturbed the programmed cell death of larval silk glands. These developmental defects correspond to loss-of-function phenotypes of BR-C Drosophila mutants in both the morphogenetic and degenerative aspects, suggesting that the critical role of BR-C in metamorphosis is evolutionarily conserved. We also demonstrate that the Sindbis virus is a useful vehicle for silencing of developmental genes in new insect models. |
| doi_str_mv | 10.1073/pnas.2136837100 |
| format | Article |
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Whether BR-C has a similar role in other holometabolous insects could not be proven without a loss-of-function genetic test, performed in a non-drosophilid species. Here we use a recombinant Sindbis virus as a tool to silence BR-C expression in the silkmoth Bombyx mori. The virus expressing a BR-C antisense RNA fragment reduced endogenous BR-C mRNA levels in infected tissues (adult wing and leg primordia) via RNA interference (RNAi). The RNAi knock-down of BR-C resulted in the failure of animals to complete the larval-pupal transition or in later morphogenetic defects, including differentiation of adult compound eyes, legs, and wings from their larval progenitors. BR-C RNAi also perturbed the programmed cell death of larval silk glands. These developmental defects correspond to loss-of-function phenotypes of BR-C Drosophila mutants in both the morphogenetic and degenerative aspects, suggesting that the critical role of BR-C in metamorphosis is evolutionarily conserved. We also demonstrate that the Sindbis virus is a useful vehicle for silencing of developmental genes in new insect models.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.2136837100</identifier><identifier>PMID: 14668449</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Adult insects ; Animals ; Base Sequence ; Biological Sciences ; Bombyx - growth & development ; Bombyx mori ; Broad-Complex protein ; Chlorocebus aethiops ; Complementary DNA ; Cricetinae ; Culicidae ; DNA Primers ; Drosophila ; Drosophila melanogaster ; Drosophila melanogaster - growth & development ; Drosophila Proteins - metabolism ; Eye - growth & development ; Insect genetics ; Insect larvae ; Larva ; Larval development ; Metamorphosis ; Metamorphosis, Biological ; Molecular Sequence Data ; Pupa ; Pupae ; RNA ; RNA, Small Interfering - pharmacology ; RNA, Small Interfering - physiology ; RNA, Viral - physiology ; Sindbis virus ; Sindbis Virus - genetics ; Transcription Factors - metabolism ; Vero Cells ; Viruses</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2003-12, Vol.100 (26), p.15607-15612</ispartof><rights>Copyright 1993-2003 National Academy of Sciences of the United States of America</rights><rights>Copyright © 2003, The National Academy of Sciences 2003</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c565t-7a2fe2faf72246cddc69f853b562f39658225e88209a613119402742fb48eb8d3</citedby><cites>FETCH-LOGICAL-c565t-7a2fe2faf72246cddc69f853b562f39658225e88209a613119402742fb48eb8d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/100/26.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/3149053$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/3149053$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27901,27902,53766,53768,57992,58225</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14668449$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Uhlirova, Mirka</creatorcontrib><creatorcontrib>Foy, Brian D.</creatorcontrib><creatorcontrib>Beaty, Barry J.</creatorcontrib><creatorcontrib>Olson, Ken E.</creatorcontrib><creatorcontrib>Riddiford, Lynn M.</creatorcontrib><creatorcontrib>Jindra, Marek</creatorcontrib><title>Use of Sindbis Virus-Mediated RNA Interference to Demonstrate a Conserved Role of Broad-Complex in Insect Metamorphosis</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>The transcription factor Broad-Complex (BR-C) is required for differentiation of adult structures as well as for the programmed death of obsolete larval organs during metamorphosis of the fruit fly Drosophila melanogaster. Whether BR-C has a similar role in other holometabolous insects could not be proven without a loss-of-function genetic test, performed in a non-drosophilid species. Here we use a recombinant Sindbis virus as a tool to silence BR-C expression in the silkmoth Bombyx mori. The virus expressing a BR-C antisense RNA fragment reduced endogenous BR-C mRNA levels in infected tissues (adult wing and leg primordia) via RNA interference (RNAi). The RNAi knock-down of BR-C resulted in the failure of animals to complete the larval-pupal transition or in later morphogenetic defects, including differentiation of adult compound eyes, legs, and wings from their larval progenitors. BR-C RNAi also perturbed the programmed cell death of larval silk glands. These developmental defects correspond to loss-of-function phenotypes of BR-C Drosophila mutants in both the morphogenetic and degenerative aspects, suggesting that the critical role of BR-C in metamorphosis is evolutionarily conserved. We also demonstrate that the Sindbis virus is a useful vehicle for silencing of developmental genes in new insect models.</description><subject>Adult insects</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological Sciences</subject><subject>Bombyx - growth & development</subject><subject>Bombyx mori</subject><subject>Broad-Complex protein</subject><subject>Chlorocebus aethiops</subject><subject>Complementary DNA</subject><subject>Cricetinae</subject><subject>Culicidae</subject><subject>DNA Primers</subject><subject>Drosophila</subject><subject>Drosophila melanogaster</subject><subject>Drosophila melanogaster - growth & development</subject><subject>Drosophila Proteins - metabolism</subject><subject>Eye - growth & development</subject><subject>Insect genetics</subject><subject>Insect larvae</subject><subject>Larva</subject><subject>Larval development</subject><subject>Metamorphosis</subject><subject>Metamorphosis, Biological</subject><subject>Molecular Sequence Data</subject><subject>Pupa</subject><subject>Pupae</subject><subject>RNA</subject><subject>RNA, Small Interfering - pharmacology</subject><subject>RNA, Small Interfering - physiology</subject><subject>RNA, Viral - physiology</subject><subject>Sindbis virus</subject><subject>Sindbis Virus - genetics</subject><subject>Transcription Factors - metabolism</subject><subject>Vero Cells</subject><subject>Viruses</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0k1v1DAQBmALgehSOHNByCckDmn9Fcc-cCjLV6UWJKBcLScZU1dJnNpOKf8eL7vqwqknW_Izoxm9Rug5JUeUNPx4nmw6YpRLxRtKyAO0okTTSgpNHqIVIayplGDiAD1J6YoQomtFHqMDKqRUQugV-nWRAAeHv_mpb33CP3xcUnUOvbcZevz18wk-nTJEBxGmDnAO-B2MYUo5FoAtXpc7xJuNDcPfVm9jsH21DuM8wC32U2mQoMv4HLIdQ5wvQ_LpKXrk7JDg2e48RBcf3n9ff6rOvnw8XZ-cVV0t61w1ljlgzrqGMSG7vu-kdqrmbS2Z41rWirEalGJEW0k5pVqUnQVzrVDQqp4fojfbvvPSjtB3MJXBBzNHP9r42wTrzf8vk780P8ON4aSRtC71r3b1MVwvkLIZfepgGOwEYUmmKYYLcT-kmsmm1qLA4y3sYkgpgrsbhhKzCdVsQjX7UEvFy3932PtdigW83oFN5b4dMUwaWkvSGLcMQ4bbXCy-xxbyYkuuUg7xznBaflXZ9g_IlcCn</recordid><startdate>20031223</startdate><enddate>20031223</enddate><creator>Uhlirova, Mirka</creator><creator>Foy, Brian D.</creator><creator>Beaty, Barry J.</creator><creator>Olson, Ken E.</creator><creator>Riddiford, Lynn M.</creator><creator>Jindra, Marek</creator><general>National Academy of Sciences</general><general>National Acad Sciences</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>7TM</scope><scope>7U9</scope><scope>H94</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20031223</creationdate><title>Use of Sindbis Virus-Mediated RNA Interference to Demonstrate a Conserved Role of Broad-Complex in Insect Metamorphosis</title><author>Uhlirova, Mirka ; Foy, Brian D. ; Beaty, Barry J. ; Olson, Ken E. ; Riddiford, Lynn M. ; Jindra, Marek</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c565t-7a2fe2faf72246cddc69f853b562f39658225e88209a613119402742fb48eb8d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Adult insects</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological Sciences</topic><topic>Bombyx - growth & development</topic><topic>Bombyx mori</topic><topic>Broad-Complex protein</topic><topic>Chlorocebus aethiops</topic><topic>Complementary DNA</topic><topic>Cricetinae</topic><topic>Culicidae</topic><topic>DNA Primers</topic><topic>Drosophila</topic><topic>Drosophila melanogaster</topic><topic>Drosophila melanogaster - growth & development</topic><topic>Drosophila Proteins - metabolism</topic><topic>Eye - growth & development</topic><topic>Insect genetics</topic><topic>Insect larvae</topic><topic>Larva</topic><topic>Larval development</topic><topic>Metamorphosis</topic><topic>Metamorphosis, Biological</topic><topic>Molecular Sequence Data</topic><topic>Pupa</topic><topic>Pupae</topic><topic>RNA</topic><topic>RNA, Small Interfering - pharmacology</topic><topic>RNA, Small Interfering - physiology</topic><topic>RNA, Viral - physiology</topic><topic>Sindbis virus</topic><topic>Sindbis Virus - genetics</topic><topic>Transcription Factors - metabolism</topic><topic>Vero Cells</topic><topic>Viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Uhlirova, Mirka</creatorcontrib><creatorcontrib>Foy, Brian D.</creatorcontrib><creatorcontrib>Beaty, Barry J.</creatorcontrib><creatorcontrib>Olson, Ken E.</creatorcontrib><creatorcontrib>Riddiford, Lynn M.</creatorcontrib><creatorcontrib>Jindra, Marek</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>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Uhlirova, Mirka</au><au>Foy, Brian D.</au><au>Beaty, Barry J.</au><au>Olson, Ken E.</au><au>Riddiford, Lynn M.</au><au>Jindra, Marek</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Use of Sindbis Virus-Mediated RNA Interference to Demonstrate a Conserved Role of Broad-Complex in Insect Metamorphosis</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>2003-12-23</date><risdate>2003</risdate><volume>100</volume><issue>26</issue><spage>15607</spage><epage>15612</epage><pages>15607-15612</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>The transcription factor Broad-Complex (BR-C) is required for differentiation of adult structures as well as for the programmed death of obsolete larval organs during metamorphosis of the fruit fly Drosophila melanogaster. Whether BR-C has a similar role in other holometabolous insects could not be proven without a loss-of-function genetic test, performed in a non-drosophilid species. Here we use a recombinant Sindbis virus as a tool to silence BR-C expression in the silkmoth Bombyx mori. The virus expressing a BR-C antisense RNA fragment reduced endogenous BR-C mRNA levels in infected tissues (adult wing and leg primordia) via RNA interference (RNAi). The RNAi knock-down of BR-C resulted in the failure of animals to complete the larval-pupal transition or in later morphogenetic defects, including differentiation of adult compound eyes, legs, and wings from their larval progenitors. BR-C RNAi also perturbed the programmed cell death of larval silk glands. These developmental defects correspond to loss-of-function phenotypes of BR-C Drosophila mutants in both the morphogenetic and degenerative aspects, suggesting that the critical role of BR-C in metamorphosis is evolutionarily conserved. We also demonstrate that the Sindbis virus is a useful vehicle for silencing of developmental genes in new insect models.</abstract><cop>United States</cop><pub>National Academy of Sciences</pub><pmid>14668449</pmid><doi>10.1073/pnas.2136837100</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 2003-12, Vol.100 (26), p.15607-15612 |
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| subjects | Adult insects Animals Base Sequence Biological Sciences Bombyx - growth & development Bombyx mori Broad-Complex protein Chlorocebus aethiops Complementary DNA Cricetinae Culicidae DNA Primers Drosophila Drosophila melanogaster Drosophila melanogaster - growth & development Drosophila Proteins - metabolism Eye - growth & development Insect genetics Insect larvae Larva Larval development Metamorphosis Metamorphosis, Biological Molecular Sequence Data Pupa Pupae RNA RNA, Small Interfering - pharmacology RNA, Small Interfering - physiology RNA, Viral - physiology Sindbis virus Sindbis Virus - genetics Transcription Factors - metabolism Vero Cells Viruses |
| title | Use of Sindbis Virus-Mediated RNA Interference to Demonstrate a Conserved Role of Broad-Complex in Insect Metamorphosis |
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