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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Targeted Mutagenesis, Precise Gene Editing, and Site-Specific Gene Insertion in Maize Using Cas9 and Guide RNA<xref ref-type="fn" rid="fn1"><sup>[OPEN]</sup></xref></title><author><name sortKey="Svitashev, Sergei" sort="Svitashev, Sergei" uniqKey="Svitashev S" first="Sergei" last="Svitashev">Sergei Svitashev</name></author><author><name sortKey="Young, Joshua K" sort="Young, Joshua K" uniqKey="Young J" first="Joshua K." last="Young">Joshua K. Young</name></author><author><name sortKey="Schwartz, Christine" sort="Schwartz, Christine" uniqKey="Schwartz C" first="Christine" last="Schwartz">Christine Schwartz</name></author><author><name sortKey="Gao, Huirong" sort="Gao, Huirong" uniqKey="Gao H" first="Huirong" last="Gao">Huirong Gao</name></author><author><name sortKey="Falco, S Carl" sort="Falco, S Carl" uniqKey="Falco S" first="S. Carl" last="Falco">S. Carl Falco</name></author><author><name sortKey="Cigan, A Mark" sort="Cigan, A Mark" uniqKey="Cigan A" first="A. Mark" last="Cigan">A. Mark Cigan</name></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">26269544</idno><idno type="pmc">4587463</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4587463</idno><idno type="RBID">PMC:4587463</idno><idno type="doi">10.1104/pp.15.00793</idno><date when="2015">2015</date><idno type="wicri:Area/Pmc/Corpus">000023</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Targeted Mutagenesis, Precise Gene Editing, and Site-Specific Gene Insertion in Maize Using Cas9 and Guide RNA<xref ref-type="fn" rid="fn1"><sup>[OPEN]</sup></xref></title><author><name sortKey="Svitashev, Sergei" sort="Svitashev, Sergei" uniqKey="Svitashev S" first="Sergei" last="Svitashev">Sergei Svitashev</name></author><author><name sortKey="Young, Joshua K" sort="Young, Joshua K" uniqKey="Young J" first="Joshua K." last="Young">Joshua K. Young</name></author><author><name sortKey="Schwartz, Christine" sort="Schwartz, Christine" uniqKey="Schwartz C" first="Christine" last="Schwartz">Christine Schwartz</name></author><author><name sortKey="Gao, Huirong" sort="Gao, Huirong" uniqKey="Gao H" first="Huirong" last="Gao">Huirong Gao</name></author><author><name sortKey="Falco, S Carl" sort="Falco, S Carl" uniqKey="Falco S" first="S. Carl" last="Falco">S. Carl Falco</name></author><author><name sortKey="Cigan, A Mark" sort="Cigan, A Mark" uniqKey="Cigan A" first="A. Mark" last="Cigan">A. Mark Cigan</name></author></analytic><series><title level="j">Plant Physiology</title><idno type="ISSN">0032-0889</idno><idno type="eISSN">1532-2548</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Genome editing and gene insertion in maize is advanced with Cas9-guide RNA technology.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Plant Physiol</journal-id><journal-id journal-id-type="iso-abbrev">Plant Physiol</journal-id><journal-id journal-id-type="hwp">plantphysiol</journal-id><journal-id journal-id-type="publisher-id">aspb</journal-id><journal-title-group><journal-title>Plant Physiology</journal-title></journal-title-group><issn pub-type="ppub">0032-0889</issn><issn pub-type="epub">1532-2548</issn><publisher><publisher-name>American Society of Plant Biologists</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26269544</article-id><article-id pub-id-type="pmc">4587463</article-id><article-id pub-id-type="publisher-id">PP201500793</article-id><article-id pub-id-type="doi">10.1104/pp.15.00793</article-id><article-categories><subj-group subj-group-type="heading"><subject>Breakthrough Technologies</subject></subj-group></article-categories><title-group><article-title>Targeted Mutagenesis, Precise Gene Editing, and Site-Specific Gene Insertion in Maize Using Cas9 and Guide RNA<xref ref-type="fn" rid="fn1"><sup>[OPEN]</sup></xref></article-title></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><name><surname>Svitashev</surname><given-names>Sergei</given-names></name><xref ref-type="author-notes" rid="afn3"><sup>1</sup></xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Young</surname><given-names>Joshua K.</given-names></name><xref ref-type="author-notes" rid="afn3"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Schwartz</surname><given-names>Christine</given-names></name></contrib><contrib contrib-type="author"><name><surname>Gao</surname><given-names>Huirong</given-names></name></contrib><contrib contrib-type="author"><name><surname>Falco</surname><given-names>S. Carl</given-names></name></contrib><contrib contrib-type="author"><name><surname>Cigan</surname><given-names>A. Mark</given-names></name><xref ref-type="corresp" rid="cor1">*</xref></contrib><aff id="aff1">Trait Enabling Technologies, DuPont Pioneer, Johnston, Iowa 50131</aff></contrib-group><author-notes><fn><p><ext-link ext-link-type="uri" xlink:href="http://www.plantphysiol.org/cgi/doi/10.1104/pp.15.00793">www.plantphysiol.org/cgi/doi/10.1104/pp.15.00793</ext-link></p></fn><fn id="afn3" fn-type="equal"><label>1</label><p>These authors contributed equally to the article.</p></fn><corresp id="cor1"><label>*</label>Address correspondence to <email>mark.cigan@pioneer.com</email>.</corresp><fn id="afn1"><p>The author responsible for distribution of materials integral to the findings presented in this article in accordance with the Journal policy described in the Instructions for Authors (<ext-link ext-link-type="uri" xlink:href="http://www.plantphysiol.org">http://www.plantphysiol.org</ext-link>) is A. Mark Cigan (<email>mark.cigan@pioneer.com</email>).</p></fn><fn id="afn2"><p>S.C.F. and A.M.C. conceived the project; S.S., J.K.Y., H.G., and A.M.C. designed the experiments; S.S., J.K.Y., and C.S. performed the experiments; S.S., J.K.Y., C.S., and A.M.C. analyzed the data; S.S. and A.M.C. wrote the article.</p></fn></author-notes><pmc-comment>Fake ppub date generated by PMC from publisher
pub-date/@pub-type='epub-ppub' </pmc-comment>
<pub-date pub-type="ppub"><month>10</month><year>2015</year></pub-date><pub-date pub-type="epub"><day>12</day><month>8</month><year>2015</year></pub-date><pub-date pub-type="pmc-release"><day>12</day><month>8</month><year>2015</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on the
<volume>169</volume><issue>2</issue><fpage>931</fpage><lpage>945</lpage><history><date date-type="received"><day>28</day><month>5</month><year>2015</year></date><date date-type="accepted"><day>07</day><month>8</month><year>2015</year></date></history><permissions><copyright-statement>© 2015 American Society of Plant Biologists. All Rights Reserved.</copyright-statement><copyright-year>2015</copyright-year></permissions><self-uri xlink:role="icon" xlink:href="PP_PP201500793_ic1.gif"></self-uri><abstract abstract-type="precis"><p>Genome editing and gene insertion in maize is advanced with Cas9-guide RNA technology.</p></abstract><abstract><p>Targeted mutagenesis, editing of endogenous maize (<italic>Zea mays</italic>) genes, and site-specific insertion of a trait gene using clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas)-guide RNA technology are reported in maize. DNA vectors expressing maize codon-optimized <italic>Streptococcus pyogenes</italic> Cas9 endonuclease and single guide RNAs were cointroduced with or without DNA repair templates into maize immature embryos by biolistic transformation targeting five different genomic regions: upstream of the <italic>liguleless1</italic> (<italic>LIG1</italic>) gene, male fertility genes (<italic>Ms26</italic> and <italic>Ms45</italic>), and acetolactate synthase (ALS) genes (<italic>ALS1</italic> and <italic>ALS2</italic>). Mutations were subsequently identified at all sites targeted, and plants containing biallelic multiplex mutations at <italic>LIG1</italic>, <italic>Ms26</italic>, and <italic>Ms45</italic> were recovered. Biolistic delivery of guide RNAs (as RNA molecules) directly into immature embryo cells containing preintegrated Cas9 also resulted in targeted mutations. Editing the <italic>ALS2</italic> gene using either single-stranded oligonucleotides or double-stranded DNA vectors as repair templates yielded chlorsulfuron-resistant plants. Double-strand breaks generated by RNA-guided Cas9 endonuclease also stimulated insertion of a trait gene at a site near <italic>LIG1</italic> by homology-directed repair. Progeny showed expected Mendelian segregation of mutations, edits, and targeted gene insertions. The examples reported in this study demonstrate the utility of Cas9-guide RNA technology as a plant genome editing tool to enhance plant breeding and crop research needed to meet growing agriculture demands of the future.</p></abstract><counts><page-count count="15"></page-count></counts><custom-meta-group><custom-meta><meta-name> DJS Export </meta-name><meta-value>v1</meta-value></custom-meta></custom-meta-group></article-meta><notes><glossary><title>Glossary</title><def-list><def-item><term id="term1">crRNA</term><def id="def1"><p>clustered regularly interspaced short palindromic repeats RNA</p></def></def-item><def-item><term id="term2">DSB</term><def id="def2"><p>double-strand break</p></def></def-item><def-item><term id="term3">gRNA</term><def id="def3"><p>guide RNA</p></def></def-item><def-item><term id="term4">HDR</term><def id="def4"><p>homology-directed repair</p></def></def-item><def-item><term id="term5">NHEJ</term><def id="def5"><p>nonhomologous end joining</p></def></def-item><def-item><term id="term6">PAM</term><def id="def6"><p>protospacer-adjacent motif</p></def></def-item><def-item><term id="term7">qPCR</term><def id="def7"><p>quantitative PCR</p></def></def-item><def-item><term id="term8">SNP</term><def id="def8"><p>single-nucleotide polymorphism</p></def></def-item><def-item><term id="term9">SSC</term><def id="def9"><p>standard sodium citrate</p></def></def-item><def-item><term id="term10">TALEN</term><def id="def10"><p>transcription activator-like effector nuclease</p></def></def-item><def-item><term id="term11">T-DNA</term><def id="def11"><p>transfer DNA</p></def></def-item><def-item><term id="term12">ZFN</term><def id="def12"><p>Zinc-finger nuclease</p></def></def-item></def-list></glossary></notes></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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