Targeted Genome Editing of Sweet Orange Using Cas9/sgRNA
Identifieur interne : 000906 ( Main/Exploration ); précédent : 000905; suivant : 000907Targeted Genome Editing of Sweet Orange Using Cas9/sgRNA
Auteurs : Hongge Jia ; Nian WangSource :
- PLoS ONE [ 1932-6203 ] ; 2014.
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : RNA, Guide.
- genetics : Citrus sinensis.
- methods : Gene Targeting, Genetic Engineering.
- Clustered Regularly Interspaced Short Palindromic Repeats.
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
Url:
DOI: 10.1371/journal.pone.0093806
PubMed: 24710347
PubMed Central: 3977896
Affiliations:
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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><idno type="wicri:Area/Pmc/Checkpoint">000694</idno><idno type="wicri:source">PubMed</idno><idno type="wicri:Area/PubMed/Corpus">000349</idno><idno type="wicri:Area/PubMed/Curation">000349</idno><idno type="wicri:Area/PubMed/Checkpoint">000349</idno><idno type="wicri:Area/Ncbi/Merge">001663</idno><idno type="wicri:Area/Ncbi/Curation">001663</idno><idno type="wicri:Area/Ncbi/Checkpoint">001663</idno><idno type="wicri:Area/Main/Merge">000906</idno><idno type="wicri:Area/Main/Curation">000906</idno><idno type="wicri:Area/Main/Exploration">000906</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><keywords scheme="KwdEn" xml:lang="en"><term>Citrus sinensis (genetics)</term><term>Clustered Regularly Interspaced Short Palindromic Repeats</term><term>Gene Targeting (methods)</term><term>Genetic Engineering (methods)</term><term>RNA, Guide (genetics)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Guide</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Citrus sinensis</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Gene Targeting</term><term>Genetic Engineering</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Clustered Regularly Interspaced Short Palindromic Repeats</term></keywords></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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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><affiliations><list></list><tree><noCountry><name sortKey="Jia, Hongge" sort="Jia, Hongge" uniqKey="Jia H" first="Hongge" last="Jia">Hongge Jia</name><name sortKey="Wang, Nian" sort="Wang, Nian" uniqKey="Wang N" first="Nian" last="Wang">Nian Wang</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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