Targeted Genome Editing of Sweet Orange Using Cas9/sgRNA
Identifieur interne : 001196 ( Pmc/Curation ); précédent : 001195; suivant : 001197Targeted Genome Editing of Sweet Orange Using Cas9/sgRNA
Auteurs : Hongge Jia ; Nian WangSource :
- PLoS ONE [ 1932-6203 ] ; 2014.
Abstract
Genetic modification, including plant breeding, has been widely used to improve crop yield and quality, as well as to increase disease resistance. Targeted genome engineering is expected to contribute significantly to future varietal improvement, and genome editing technologies using zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9/single guide RNA (sgRNA) have already been successfully used to genetically modify plants. However, to date, there has been no reported use of any of the current genome editing approaches in sweet orange, an important fruit crop. In this study, we first developed a novel tool, Xcc-facilitated agroinfiltration, for enhancing transient protein expression in sweet orange leaves. We then successfully employed Xcc-facilitated agroinfiltration to deliver Cas9, along with a synthetic sgRNA targeting the
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DOI: 10.1371/journal.pone.0093806
PubMed: 24710347
PubMed Central: 3977896
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Targeted Genome Editing of Sweet Orange Using Cas9/sgRNA</title><author><name sortKey="Jia, Hongge" sort="Jia, Hongge" uniqKey="Jia H" first="Hongge" last="Jia">Hongge Jia</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Wang, Nian" sort="Wang, Nian" uniqKey="Wang N" first="Nian" last="Wang">Nian Wang</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">24710347</idno><idno type="pmc">3977896</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3977896</idno><idno type="RBID">PMC:3977896</idno><idno type="doi">10.1371/journal.pone.0093806</idno><date when="2014">2014</date><idno type="wicri:Area/Pmc/Corpus">001197</idno><idno type="wicri:Area/Pmc/Curation">001196</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Targeted Genome Editing of Sweet Orange Using Cas9/sgRNA</title><author><name sortKey="Jia, Hongge" sort="Jia, Hongge" uniqKey="Jia H" first="Hongge" last="Jia">Hongge Jia</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Wang, Nian" sort="Wang, Nian" uniqKey="Wang N" first="Nian" last="Wang">Nian Wang</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Genetic modification, including plant breeding, has been widely used to improve crop yield and quality, as well as to increase disease resistance. Targeted genome engineering is expected to contribute significantly to future varietal improvement, and genome editing technologies using zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9/single guide RNA (sgRNA) have already been successfully used to genetically modify plants. However, to date, there has been no reported use of any of the current genome editing approaches in sweet orange, an important fruit crop. In this study, we first developed a novel tool, Xcc-facilitated agroinfiltration, for enhancing transient protein expression in sweet orange leaves. We then successfully employed Xcc-facilitated agroinfiltration to deliver Cas9, along with a synthetic sgRNA targeting the <italic>CsPDS</italic> gene, into sweet orange. DNA sequencing confirmed that the <italic>CsPDS</italic> gene was mutated at the target site in treated sweet orange leaves. The mutation rate using the Cas9/sgRNA system was approximately 3.2 to 3.9%. Off-target mutagenesis was not detected for <italic>CsPDS</italic>-related DNA sequences in our study. This is the first report of targeted genome modification in citrus using the Cas9/sgRNA system—a system that holds significant promise for the study of citrus gene function and for targeted genetic modification.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Curtin, Sj" uniqKey="Curtin S">SJ Curtin</name></author><author><name sortKey="Anderson, Je" uniqKey="Anderson J">JE Anderson</name></author><author><name sortKey="Starker, Cg" uniqKey="Starker C">CG Starker</name></author><author><name sortKey="Baltes, Nj" uniqKey="Baltes N">NJ Baltes</name></author><author><name sortKey="Mani, D" uniqKey="Mani D">D Mani</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="De Pater, S" uniqKey="De Pater S">S de Pater</name></author><author><name sortKey="Pinas, Je" uniqKey="Pinas J">JE Pinas</name></author><author><name sortKey="Hooykaas, Pj" uniqKey="Hooykaas P">PJ 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sortKey="Carroll, D" uniqKey="Carroll D">D Carroll</name></author><author><name sortKey="Drews, Gn" uniqKey="Drews G">GN Drews</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shukla, Vk" uniqKey="Shukla V">VK Shukla</name></author><author><name sortKey="Doyon, Y" uniqKey="Doyon Y">Y Doyon</name></author><author><name sortKey="Miller, Jc" uniqKey="Miller J">JC Miller</name></author><author><name sortKey="Dekelver, Rc" uniqKey="Dekelver R">RC DeKelver</name></author><author><name sortKey="Moehle, Ea" uniqKey="Moehle E">EA Moehle</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Townsend, Ja" uniqKey="Townsend J">JA Townsend</name></author><author><name sortKey="Wright, Da" uniqKey="Wright D">DA Wright</name></author><author><name sortKey="Winfrey, Rj" uniqKey="Winfrey R">RJ Winfrey</name></author><author><name sortKey="Fu, F" uniqKey="Fu F">F Fu</name></author><author><name sortKey="Maeder, Ml" uniqKey="Maeder M">ML Maeder</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gaj, T" uniqKey="Gaj T">T Gaj</name></author><author><name sortKey="Gersbach, Ca" uniqKey="Gersbach C">CA Gersbach</name></author><author><name sortKey="Barbas, Cf" uniqKey="Barbas C">CF Barbas</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chen, K" uniqKey="Chen K">K Chen</name></author><author><name sortKey="Gao, C" uniqKey="Gao C">C Gao</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Boch, J" uniqKey="Boch J">J Boch</name></author><author><name sortKey="Scholze, H" uniqKey="Scholze H">H Scholze</name></author><author><name sortKey="Schornack, S" uniqKey="Schornack S">S Schornack</name></author><author><name sortKey="Landgraf, A" uniqKey="Landgraf A">A Landgraf</name></author><author><name sortKey="Hahn, S" uniqKey="Hahn S">S Hahn</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Moscou, Mj" uniqKey="Moscou M">MJ Moscou</name></author><author><name sortKey="Bogdanove, Aj" uniqKey="Bogdanove A">AJ Bogdanove</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cermak, T" uniqKey="Cermak T">T Cermak</name></author><author><name sortKey="Doyle, El" uniqKey="Doyle E">EL Doyle</name></author><author><name sortKey="Christian, M" uniqKey="Christian M">M Christian</name></author><author><name sortKey="Wang, L" uniqKey="Wang L">L Wang</name></author><author><name sortKey="Zhang, Y" uniqKey="Zhang Y">Y Zhang</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Li, T" uniqKey="Li T">T Li</name></author><author><name sortKey="Liu, B" uniqKey="Liu B">B Liu</name></author><author><name sortKey="Spalding, Mh" uniqKey="Spalding M">MH Spalding</name></author><author><name sortKey="Weeks, Dp" uniqKey="Weeks D">DP Weeks</name></author><author><name sortKey="Yang, B" uniqKey="Yang B">B Yang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zhang, Y" uniqKey="Zhang Y">Y Zhang</name></author><author><name sortKey="Zhang, F" uniqKey="Zhang F">F Zhang</name></author><author><name sortKey="Li, X" uniqKey="Li X">X Li</name></author><author><name sortKey="Baller, Ja" uniqKey="Baller J">JA Baller</name></author><author><name sortKey="Qi, Y" uniqKey="Qi Y">Y Qi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wendt, T" uniqKey="Wendt T">T Wendt</name></author><author><name sortKey="Holm, Pb" uniqKey="Holm P">PB Holm</name></author><author><name sortKey="Starker, Cg" uniqKey="Starker C">CG Starker</name></author><author><name sortKey="Christian, M" uniqKey="Christian M">M Christian</name></author><author><name sortKey="Voytas, Df" uniqKey="Voytas D">DF Voytas</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Wiedenheft, B" uniqKey="Wiedenheft B">B 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Ma</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mali, P" uniqKey="Mali P">P Mali</name></author><author><name sortKey="Yang, L" uniqKey="Yang L">L Yang</name></author><author><name sortKey="Esvelt, Km" uniqKey="Esvelt K">KM Esvelt</name></author><author><name sortKey="Aach, J" uniqKey="Aach J">J Aach</name></author><author><name sortKey="Guell, M" uniqKey="Guell M">M Guell</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Strau, A" uniqKey="Strau A">A Strauβ</name></author><author><name sortKey="Lahaye, T" uniqKey="Lahaye T">T Lahaye</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Streubel, J" uniqKey="Streubel J">J Streubel</name></author><author><name sortKey="Blucher, C" uniqKey="Blucher C">C Blücher</name></author><author><name sortKey="Landgraf, A" uniqKey="Landgraf A">A Landgraf</name></author><author><name sortKey="Boch, J" uniqKey="Boch J">J Boch</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Nekrasov, V" uniqKey="Nekrasov V">V Nekrasov</name></author><author><name sortKey="Staskawicz, B" uniqKey="Staskawicz B">B Staskawicz</name></author><author><name sortKey="Weigel, D" uniqKey="Weigel D">D Weigel</name></author><author><name sortKey="Jones, Jd" uniqKey="Jones J">JD Jones</name></author><author><name sortKey="Kamoun, S" uniqKey="Kamoun S">S Kamoun</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Xie, K" uniqKey="Xie K">K Xie</name></author><author><name sortKey="Yang, Y" uniqKey="Yang Y">Y Yang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shan, Q" uniqKey="Shan Q">Q Shan</name></author><author><name sortKey="Wang, Y" uniqKey="Wang Y">Y Wang</name></author><author><name sortKey="Li, J" uniqKey="Li J">J Li</name></author><author><name sortKey="Zhang, Y" uniqKey="Zhang Y">Y Zhang</name></author><author><name sortKey="Chen, K" uniqKey="Chen K">K Chen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Xu, Q" uniqKey="Xu Q">Q Xu</name></author><author><name sortKey="Chen, Ll" uniqKey="Chen L">LL Chen</name></author><author><name sortKey="Ruan, X" uniqKey="Ruan X">X Ruan</name></author><author><name sortKey="Chen, D" uniqKey="Chen D">D Chen</name></author><author><name sortKey="Zhu, A" uniqKey="Zhu A">A Zhu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Talon, M" uniqKey="Talon M">M Talon</name></author><author><name sortKey="Gmitter, Fg" uniqKey="Gmitter F">FG Gmitter</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jia, H" uniqKey="Jia H">H Jia</name></author><author><name sortKey="Liao, M" uniqKey="Liao M">M Liao</name></author><author><name sortKey="Verbelen, Jp" uniqKey="Verbelen J">JP Verbelen</name></author><author><name sortKey="Vissenberg, K" uniqKey="Vissenberg K">K Vissenberg</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jia, H" uniqKey="Jia H">H Jia</name></author><author><name sortKey="Pang, Y" uniqKey="Pang Y">Y Pang</name></author><author><name sortKey="Chen, X" uniqKey="Chen X">X Chen</name></author><author><name sortKey="Fang, R" uniqKey="Fang R">R Fang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Liu, Z" uniqKey="Liu Z">Z Liu</name></author><author><name sortKey="Wang, J" uniqKey="Wang J">J Wang</name></author><author><name sortKey="Huang, X" uniqKey="Huang X">X Huang</name></author><author><name sortKey="Xu, W" uniqKey="Xu W">W Xu</name></author><author><name sortKey="Liu, Z" uniqKey="Liu Z">Z Liu</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Swarup, S" uniqKey="Swarup S">S Swarup</name></author><author><name sortKey="Yang, Y" uniqKey="Yang Y">Y Yang</name></author><author><name sortKey="Kingsley, M" uniqKey="Kingsley M">M Kingsley</name></author><author><name sortKey="Gabriel, D" uniqKey="Gabriel D">D Gabriel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yan, Q" uniqKey="Yan Q">Q Yan</name></author><author><name sortKey="Wang, N" uniqKey="Wang N">N Wang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Brunings, Am" uniqKey="Brunings A">AM Brunings</name></author><author><name sortKey="Gabriel, Dw" uniqKey="Gabriel D">DW Gabriel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Li, Jf" uniqKey="Li J">JF Li</name></author><author><name sortKey="Norville, Je" uniqKey="Norville J">JE Norville</name></author><author><name sortKey="Aach, J" uniqKey="Aach J">J Aach</name></author><author><name sortKey="Mccormack, M" uniqKey="Mccormack M">M McCormack</name></author><author><name sortKey="Zhang, D" uniqKey="Zhang D">D Zhang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ding, Sw" uniqKey="Ding S">SW Ding</name></author><author><name sortKey="Rathjen, Jp" uniqKey="Rathjen J">JP Rathjen</name></author><author><name sortKey="Li, Wx" uniqKey="Li W">WX Li</name></author><author><name sortKey="Swanson, R" uniqKey="Swanson R">R Swanson</name></author><author><name sortKey="Healy, H" uniqKey="Healy H">H Healy</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fu, Y" uniqKey="Fu Y">Y Fu</name></author><author><name sortKey="Foden, Ja" uniqKey="Foden J">JA Foden</name></author><author><name sortKey="Khayter, C" uniqKey="Khayter C">C Khayter</name></author><author><name sortKey="Maeder, Ml" uniqKey="Maeder M">ML Maeder</name></author><author><name sortKey="Reyon, D" uniqKey="Reyon D">D Reyon</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Belhai, K" uniqKey="Belhai K">K Belhai</name></author><author><name sortKey="Chaparro Garcia, A" uniqKey="Chaparro Garcia A">A Chaparro-Garcia</name></author><author><name sortKey="Kamoun, S" uniqKey="Kamoun S">S Kamoun</name></author><author><name sortKey="Nekrasov, V" uniqKey="Nekrasov V">V Nekrasov</name></author></analytic></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">PLoS One</journal-id><journal-id journal-id-type="iso-abbrev">PLoS ONE</journal-id><journal-id journal-id-type="publisher-id">plos</journal-id><journal-id journal-id-type="pmc">plosone</journal-id><journal-title-group><journal-title>PLoS ONE</journal-title></journal-title-group><issn pub-type="epub">1932-6203</issn><publisher><publisher-name>Public Library of Science</publisher-name><publisher-loc>San Francisco, USA</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">24710347</article-id><article-id pub-id-type="pmc">3977896</article-id><article-id pub-id-type="publisher-id">PONE-D-13-52039</article-id><article-id pub-id-type="doi">10.1371/journal.pone.0093806</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Biology and Life Sciences</subject><subj-group><subject>Agriculture</subject><subj-group><subject>Agricultural Biotechnology</subject><subj-group><subject>Genetically Modified Organisms</subject><subj-group><subject>Transgenic Plants</subject></subj-group></subj-group></subj-group><subj-group><subject>Agronomy</subject><subj-group><subject>Plant Breeding</subject></subj-group></subj-group><subj-group><subject>Crops</subject></subj-group></subj-group><subj-group><subject>Biotechnology</subject><subj-group><subject>Plant Biotechnology</subject><subj-group><subject>Plant Genomics</subject></subj-group></subj-group></subj-group><subj-group><subject>Genetics</subject><subj-group><subject>Genomics</subject><subj-group><subject>Functional Genomics</subject></subj-group></subj-group><subj-group><subject>Plant Genetics</subject><subj-group><subject>Crop Genetics</subject></subj-group></subj-group></subj-group><subj-group><subject>Organisms</subject><subj-group><subject>Plants</subject></subj-group></subj-group><subj-group><subject>Plant Science</subject></subj-group></subj-group></article-categories><title-group><article-title>Targeted Genome Editing of Sweet Orange Using Cas9/sgRNA</article-title><alt-title alt-title-type="running-head">Genome Editing of Sweet Orange</alt-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Jia</surname><given-names>Hongge</given-names></name><xref ref-type="aff" rid="aff1"></xref></contrib><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Nian</given-names></name><xref ref-type="aff" rid="aff1"></xref><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib></contrib-group><aff id="aff1"><addr-line>Citrus Research and Education Center, Department of Microbiology and Cell Science, University of Florida, Lake Alfred, Florida, United States of America</addr-line></aff><contrib-group><contrib contrib-type="editor"><name><surname>Prasad</surname><given-names>Manoj</given-names></name><role>Editor</role><xref ref-type="aff" rid="edit1"></xref></contrib></contrib-group><aff id="edit1"><addr-line>National Institute of Plant Genome Research, India</addr-line></aff><author-notes><corresp id="cor1">* E-mail: <email>nianwang@ufl.edu</email></corresp><fn fn-type="conflict"><p><bold>Competing Interests: </bold>The authors have declared that no competing interests exist.</p></fn><fn fn-type="con"><p>Conceived and designed the experiments: NW HGJ. Performed the experiments: HGJ. Analyzed the data: NW HGJ. Wrote the paper: NW HGJ.</p></fn></author-notes><pub-date pub-type="collection"><year>2014</year></pub-date><pub-date pub-type="epub"><day>7</day><month>4</month><year>2014</year></pub-date><volume>9</volume><issue>4</issue><elocation-id>e93806</elocation-id><history><date date-type="received"><day>11</day><month>12</month><year>2013</year></date><date date-type="accepted"><day>6</day><month>3</month><year>2014</year></date></history><permissions><copyright-year>2014</copyright-year><copyright-holder>Jia, Wang</copyright-holder><license><license-p>This is an open-access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p></license></permissions><abstract><p>Genetic modification, including plant breeding, has been widely used to improve crop yield and quality, as well as to increase disease resistance. Targeted genome engineering is expected to contribute significantly to future varietal improvement, and genome editing technologies using zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9/single guide RNA (sgRNA) have already been successfully used to genetically modify plants. However, to date, there has been no reported use of any of the current genome editing approaches in sweet orange, an important fruit crop. In this study, we first developed a novel tool, Xcc-facilitated agroinfiltration, for enhancing transient protein expression in sweet orange leaves. We then successfully employed Xcc-facilitated agroinfiltration to deliver Cas9, along with a synthetic sgRNA targeting the <italic>CsPDS</italic> gene, into sweet orange. DNA sequencing confirmed that the <italic>CsPDS</italic> gene was mutated at the target site in treated sweet orange leaves. The mutation rate using the Cas9/sgRNA system was approximately 3.2 to 3.9%. Off-target mutagenesis was not detected for <italic>CsPDS</italic>-related DNA sequences in our study. This is the first report of targeted genome modification in citrus using the Cas9/sgRNA system—a system that holds significant promise for the study of citrus gene function and for targeted genetic modification.</p></abstract><funding-group><funding-statement>This work was supported by the Citrus Research and Development Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.</funding-statement></funding-group><counts><page-count count="6"></page-count></counts></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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