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In vivo functional genomic studies of sterol carrier protein-2 gene in the yellow fever mosquito
A simple and efficient DNA delivery method to introduce extrachromosomal DNA into mosquito embryos would significantly aid functional genomic studies. The conventional method for delivery of DNA into insects is to inject the DNA directly into the embryos. Taking advantage of the unique aspects of mo...
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| Published in: | PloS one 2011-03, Vol.6 (3), p.e18030-e18030 |
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| creator | Peng, Rong Maklokova, Vilena I Chandrashekhar, Jayadevi H Lan, Que |
| description | A simple and efficient DNA delivery method to introduce extrachromosomal DNA into mosquito embryos would significantly aid functional genomic studies. The conventional method for delivery of DNA into insects is to inject the DNA directly into the embryos. Taking advantage of the unique aspects of mosquito reproductive physiology during vitellogenesis and an in vivo transfection reagent that mediates DNA uptake in cells via endocytosis, we have developed a new method to introduce DNA into mosquito embryos vertically via microinjection of DNA vectors in vitellogenic females without directly manipulating the embryos. Our method was able to introduce inducible gene expression vectors transiently into F0 mosquitoes to perform functional studies in vivo without transgenic lines. The high efficiency of expression knockdown was reproducible with more than 70% of the F0 individuals showed sufficient gene expression suppression ( |
| doi_str_mv | 10.1371/journal.pone.0018030 |
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The conventional method for delivery of DNA into insects is to inject the DNA directly into the embryos. Taking advantage of the unique aspects of mosquito reproductive physiology during vitellogenesis and an in vivo transfection reagent that mediates DNA uptake in cells via endocytosis, we have developed a new method to introduce DNA into mosquito embryos vertically via microinjection of DNA vectors in vitellogenic females without directly manipulating the embryos. Our method was able to introduce inducible gene expression vectors transiently into F0 mosquitoes to perform functional studies in vivo without transgenic lines. The high efficiency of expression knockdown was reproducible with more than 70% of the F0 individuals showed sufficient gene expression suppression (<30% of the controls' levels). At the cohort level, AeSCP-2 expression knockdown in early instar larvae resulted in detectable phenotypes of the expression deficiency such as high mortality, lowered fertility, and distorted sex ratio after induction of AeSCP-2 siRNA expression in vivo. The results further confirmed the important role of AeSCP-2 in the development and reproduction of A. aegypti. In this study, we proved that extrachromosomal transient expression of an inducible gene from a DNA vector vertically delivered via vitellogenic females can be used to manipulate gene expression in F0 generation. This new method will be a simple and efficient tool for in vivo functional genomic studies in mosquitoes.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0018030</identifier><identifier>PMID: 21437205</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Aedes ; Aedes - genetics ; Aedes - growth & development ; Aedes aegypti ; Amino acids ; Animals ; Apis mellifera ; Aquatic insects ; beta-Galactosidase - metabolism ; Biology ; Carrier Proteins - genetics ; Carrier Proteins - metabolism ; Culicidae ; Deoxyribonucleic acid ; Diptera ; DNA ; DNA - genetics ; Domestication ; Drosophila melanogaster ; Eggs ; Embryos ; Endocytosis ; Expression vectors ; Extrachromosomal DNA ; Female ; Females ; Fertility ; Fever ; Gene expression ; Gene Expression Regulation ; Gene Knockdown Techniques ; Gene Transfer Techniques ; Genes ; Genes, Insect - genetics ; Genetic aspects ; Genetic engineering ; Genetic research ; Genetic vectors ; Genetic Vectors - genetics ; Genomics ; Genomics - methods ; Health aspects ; Heat-Shock Response - genetics ; In vivo methods and tests ; Insect Proteins - genetics ; Insect Proteins - metabolism ; Insects ; Laboratories ; Larva - metabolism ; Larvae ; Metabolism ; Microinjection ; Mosquitoes ; Mutation ; Oocytes - cytology ; Oocytes - metabolism ; Proteins ; Reproduction ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; RNA, Small Interfering - metabolism ; Sex ratio ; siRNA ; Stegomyia ; Steroids (Organic compounds) ; Sterols ; Transfection ; Vector-borne diseases ; Vitellogenesis ; Yellow fever ; Yellow Fever - genetics ; Yellow Fever - parasitology</subject><ispartof>PloS one, 2011-03, Vol.6 (3), p.e18030-e18030</ispartof><rights>COPYRIGHT 2011 Public Library of Science</rights><rights>2011 Peng et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>Peng et al. 2011</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c723t-beadff5032c30905114b841ce295ae8f5197339e46d4f084e9957307f93b09ba3</citedby><cites>FETCH-LOGICAL-c723t-beadff5032c30905114b841ce295ae8f5197339e46d4f084e9957307f93b09ba3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/1292421407/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/1292421407?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25752,27923,27924,37011,37012,44589,53790,53792,74897</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21437205$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Hansen, Immo</contributor><creatorcontrib>Peng, Rong</creatorcontrib><creatorcontrib>Maklokova, Vilena I</creatorcontrib><creatorcontrib>Chandrashekhar, Jayadevi H</creatorcontrib><creatorcontrib>Lan, Que</creatorcontrib><title>In vivo functional genomic studies of sterol carrier protein-2 gene in the yellow fever mosquito</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>A simple and efficient DNA delivery method to introduce extrachromosomal DNA into mosquito embryos would significantly aid functional genomic studies. The conventional method for delivery of DNA into insects is to inject the DNA directly into the embryos. Taking advantage of the unique aspects of mosquito reproductive physiology during vitellogenesis and an in vivo transfection reagent that mediates DNA uptake in cells via endocytosis, we have developed a new method to introduce DNA into mosquito embryos vertically via microinjection of DNA vectors in vitellogenic females without directly manipulating the embryos. Our method was able to introduce inducible gene expression vectors transiently into F0 mosquitoes to perform functional studies in vivo without transgenic lines. The high efficiency of expression knockdown was reproducible with more than 70% of the F0 individuals showed sufficient gene expression suppression (<30% of the controls' levels). At the cohort level, AeSCP-2 expression knockdown in early instar larvae resulted in detectable phenotypes of the expression deficiency such as high mortality, lowered fertility, and distorted sex ratio after induction of AeSCP-2 siRNA expression in vivo. The results further confirmed the important role of AeSCP-2 in the development and reproduction of A. aegypti. In this study, we proved that extrachromosomal transient expression of an inducible gene from a DNA vector vertically delivered via vitellogenic females can be used to manipulate gene expression in F0 generation. This new method will be a simple and efficient tool for in vivo functional genomic studies in mosquitoes.</description><subject>Aedes</subject><subject>Aedes - genetics</subject><subject>Aedes - growth & development</subject><subject>Aedes aegypti</subject><subject>Amino acids</subject><subject>Animals</subject><subject>Apis mellifera</subject><subject>Aquatic insects</subject><subject>beta-Galactosidase - metabolism</subject><subject>Biology</subject><subject>Carrier Proteins - genetics</subject><subject>Carrier Proteins - metabolism</subject><subject>Culicidae</subject><subject>Deoxyribonucleic acid</subject><subject>Diptera</subject><subject>DNA</subject><subject>DNA - genetics</subject><subject>Domestication</subject><subject>Drosophila melanogaster</subject><subject>Eggs</subject><subject>Embryos</subject><subject>Endocytosis</subject><subject>Expression vectors</subject><subject>Extrachromosomal DNA</subject><subject>Female</subject><subject>Females</subject><subject>Fertility</subject><subject>Fever</subject><subject>Gene expression</subject><subject>Gene Expression Regulation</subject><subject>Gene Knockdown Techniques</subject><subject>Gene Transfer Techniques</subject><subject>Genes</subject><subject>Genes, Insect - genetics</subject><subject>Genetic aspects</subject><subject>Genetic engineering</subject><subject>Genetic research</subject><subject>Genetic vectors</subject><subject>Genetic Vectors - genetics</subject><subject>Genomics</subject><subject>Genomics - methods</subject><subject>Health aspects</subject><subject>Heat-Shock Response - genetics</subject><subject>In vivo methods and tests</subject><subject>Insect Proteins - genetics</subject><subject>Insect Proteins - metabolism</subject><subject>Insects</subject><subject>Laboratories</subject><subject>Larva - metabolism</subject><subject>Larvae</subject><subject>Metabolism</subject><subject>Microinjection</subject><subject>Mosquitoes</subject><subject>Mutation</subject><subject>Oocytes - cytology</subject><subject>Oocytes - metabolism</subject><subject>Proteins</subject><subject>Reproduction</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>RNA, Small Interfering - metabolism</subject><subject>Sex ratio</subject><subject>siRNA</subject><subject>Stegomyia</subject><subject>Steroids (Organic compounds)</subject><subject>Sterols</subject><subject>Transfection</subject><subject>Vector-borne diseases</subject><subject>Vitellogenesis</subject><subject>Yellow fever</subject><subject>Yellow Fever - genetics</subject><subject>Yellow Fever - parasitology</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNqNk12L1DAUhoso7jr6D0QLguLFjPlok-ZmYVn8GFhY8Os2punJTIZOM5uko_vvTZ3uMpUFJRcJyXPeJO85J8ueY7TAlON3G9f7TrWLnetggRCuEEUPslMsKJkzgujDo_VJ9iSEDUIlrRh7nJ0QXFBOUHma_Vh2-d7uXW76TkfrkmK-gs5trc5D7BsLIXcmLcG7NtfKews-33kXwXZzMrCQ2y6Pa8hvoG3dz9zAPiFbF657G93T7JFRbYBn4zzLvn14__Xi0_zy6uPy4vxyrjmhcV6DaowpESWaIoFKjIu6KrAGIkoFlSmx4JQKKFhTGFQVIETJKeJG0BqJWtFZ9vKgu2tdkKM5QWIiSJG-i3gilgeicWojd95ulb-RTln5Z8P5lVQ-Wt2CVFXD6lqYpmrKQtdaIcMrRjg3jLGqHrTOxtv6eguNhi561U5EpyedXcuV20uKGBKkTAJvRgHvrnsIUW5t0MlA1YHrg6xS6hipmPg3WVY85Z6xRL76i7zfhpFaqfRT2xmXHqgHTXlecCZSKSWnZ9niHiqNBlJppJIzNu1PAt5OAhIT4VdcqT4Eufzy-f_Zq-9T9vURuwbVxnVwbT_UapiCxQHU3oXgwdxlAyM5dMytG3LoGDl2TAp7cZzJu6DbFqG_Ab_4EBE</recordid><startdate>20110318</startdate><enddate>20110318</enddate><creator>Peng, Rong</creator><creator>Maklokova, Vilena I</creator><creator>Chandrashekhar, Jayadevi H</creator><creator>Lan, Que</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>F1W</scope><scope>H95</scope><scope>H97</scope><scope>L.G</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20110318</creationdate><title>In vivo functional genomic studies of sterol carrier protein-2 gene in the yellow fever mosquito</title><author>Peng, Rong ; Maklokova, Vilena I ; Chandrashekhar, Jayadevi H ; Lan, Que</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c723t-beadff5032c30905114b841ce295ae8f5197339e46d4f084e9957307f93b09ba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Aedes</topic><topic>Aedes - genetics</topic><topic>Aedes - growth & development</topic><topic>Aedes aegypti</topic><topic>Amino acids</topic><topic>Animals</topic><topic>Apis mellifera</topic><topic>Aquatic insects</topic><topic>beta-Galactosidase - metabolism</topic><topic>Biology</topic><topic>Carrier Proteins - genetics</topic><topic>Carrier Proteins - metabolism</topic><topic>Culicidae</topic><topic>Deoxyribonucleic acid</topic><topic>Diptera</topic><topic>DNA</topic><topic>DNA - genetics</topic><topic>Domestication</topic><topic>Drosophila melanogaster</topic><topic>Eggs</topic><topic>Embryos</topic><topic>Endocytosis</topic><topic>Expression vectors</topic><topic>Extrachromosomal DNA</topic><topic>Female</topic><topic>Females</topic><topic>Fertility</topic><topic>Fever</topic><topic>Gene expression</topic><topic>Gene Expression Regulation</topic><topic>Gene Knockdown Techniques</topic><topic>Gene Transfer Techniques</topic><topic>Genes</topic><topic>Genes, Insect - genetics</topic><topic>Genetic aspects</topic><topic>Genetic engineering</topic><topic>Genetic research</topic><topic>Genetic vectors</topic><topic>Genetic Vectors - 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The conventional method for delivery of DNA into insects is to inject the DNA directly into the embryos. Taking advantage of the unique aspects of mosquito reproductive physiology during vitellogenesis and an in vivo transfection reagent that mediates DNA uptake in cells via endocytosis, we have developed a new method to introduce DNA into mosquito embryos vertically via microinjection of DNA vectors in vitellogenic females without directly manipulating the embryos. Our method was able to introduce inducible gene expression vectors transiently into F0 mosquitoes to perform functional studies in vivo without transgenic lines. The high efficiency of expression knockdown was reproducible with more than 70% of the F0 individuals showed sufficient gene expression suppression (<30% of the controls' levels). At the cohort level, AeSCP-2 expression knockdown in early instar larvae resulted in detectable phenotypes of the expression deficiency such as high mortality, lowered fertility, and distorted sex ratio after induction of AeSCP-2 siRNA expression in vivo. The results further confirmed the important role of AeSCP-2 in the development and reproduction of A. aegypti. In this study, we proved that extrachromosomal transient expression of an inducible gene from a DNA vector vertically delivered via vitellogenic females can be used to manipulate gene expression in F0 generation. This new method will be a simple and efficient tool for in vivo functional genomic studies in mosquitoes.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>21437205</pmid><doi>10.1371/journal.pone.0018030</doi><tpages>e18030</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2011-03, Vol.6 (3), p.e18030-e18030 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1292421407 |
| source | Publicly Available Content Database; PubMed Central (Training) |
| subjects | Aedes Aedes - genetics Aedes - growth & development Aedes aegypti Amino acids Animals Apis mellifera Aquatic insects beta-Galactosidase - metabolism Biology Carrier Proteins - genetics Carrier Proteins - metabolism Culicidae Deoxyribonucleic acid Diptera DNA DNA - genetics Domestication Drosophila melanogaster Eggs Embryos Endocytosis Expression vectors Extrachromosomal DNA Female Females Fertility Fever Gene expression Gene Expression Regulation Gene Knockdown Techniques Gene Transfer Techniques Genes Genes, Insect - genetics Genetic aspects Genetic engineering Genetic research Genetic vectors Genetic Vectors - genetics Genomics Genomics - methods Health aspects Heat-Shock Response - genetics In vivo methods and tests Insect Proteins - genetics Insect Proteins - metabolism Insects Laboratories Larva - metabolism Larvae Metabolism Microinjection Mosquitoes Mutation Oocytes - cytology Oocytes - metabolism Proteins Reproduction RNA, Messenger - genetics RNA, Messenger - metabolism RNA, Small Interfering - metabolism Sex ratio siRNA Stegomyia Steroids (Organic compounds) Sterols Transfection Vector-borne diseases Vitellogenesis Yellow fever Yellow Fever - genetics Yellow Fever - parasitology |
| title | In vivo functional genomic studies of sterol carrier protein-2 gene in the yellow fever mosquito |
| url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-10T09%3A07%3A44IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=In%20vivo%20functional%20genomic%20studies%20of%20sterol%20carrier%20protein-2%20gene%20in%20the%20yellow%20fever%20mosquito&rft.jtitle=PloS%20one&rft.au=Peng,%20Rong&rft.date=2011-03-18&rft.volume=6&rft.issue=3&rft.spage=e18030&rft.epage=e18030&rft.pages=e18030-e18030&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0018030&rft_dat=%3Cgale_plos_%3EA476900173%3C/gale_plos_%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c723t-beadff5032c30905114b841ce295ae8f5197339e46d4f084e9957307f93b09ba3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=1292421407&rft_id=info:pmid/21437205&rft_galeid=A476900173&rfr_iscdi=true |