Manipulating the Biosynthesis of Bioactive Compound Alkaloids for Next-Generation Metabolic Engineering in Opium Poppy Using CRISPR-Cas 9 Genome Editing Technology
Identifieur interne : 000139 ( Main/Exploration ); précédent : 000138; suivant : 000140Manipulating the Biosynthesis of Bioactive Compound Alkaloids for Next-Generation Metabolic Engineering in Opium Poppy Using CRISPR-Cas 9 Genome Editing Technology
Auteurs : Yagiz Alagoz [Turquie, Australie] ; Tugba Gurkok [Turquie] ; Baohong Zhang [États-Unis] ; Turgay Unver [Turquie]Source :
- Scientific Reports [ 2045-2322 ] ; 2016.
Abstract
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated9 (Cas9) endonuclease system is a powerful RNA-guided genome editing tool. CRISPR/Cas9 has been well studied in model plant species for targeted genome editing. However, few studies have been reported on plant species without whole genome sequence information. Currently, no study has been performed to manipulate metabolic pathways using CRISPR/Cas9. In this study, the type II CRISPR/SpCas9 system was used to knock out, via nonhomologous end-joining genome repair, the
Url:
DOI: 10.1038/srep30910
PubMed: 27483984
PubMed Central: 4971470
Affiliations:
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Manipulating the Biosynthesis of Bioactive Compound Alkaloids for Next-Generation Metabolic Engineering in Opium Poppy Using CRISPR-Cas 9 Genome Editing Technology</title><author><name sortKey="Alagoz, Yagiz" sort="Alagoz, Yagiz" uniqKey="Alagoz Y" first="Yagiz" last="Alagoz">Yagiz Alagoz</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Biology, Faculty of Science, Çankırı Karatekin University</institution>, Çankırı18100,<country>Turkey</country></nlm:aff><country xml:lang="fr">Turquie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Hawkesbury Institute for the Environment, Western Sydney University</institution>, Richmond, New South Wales 2753,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Gurkok, Tugba" sort="Gurkok, Tugba" uniqKey="Gurkok T" first="Tugba" last="Gurkok">Tugba Gurkok</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Biology, Faculty of Science, Çankırı Karatekin University</institution>, Çankırı18100,<country>Turkey</country></nlm:aff><country xml:lang="fr">Turquie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Zhang, Baohong" sort="Zhang, Baohong" uniqKey="Zhang B" first="Baohong" last="Zhang">Baohong Zhang</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Department of Biology, East Carolina University</institution>, Greenville, NC 27858,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Unver, Turgay" sort="Unver, Turgay" uniqKey="Unver T" first="Turgay" last="Unver">Turgay Unver</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Biology, Faculty of Science, Çankırı Karatekin University</institution>, Çankırı18100,<country>Turkey</country></nlm:aff><country xml:lang="fr">Turquie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Izmir International Biomedicine and Genome Institute (iBG-izmir), Dokuz Eylul University</institution>, Balcova 35340 Izmir,<country>Turkey</country></nlm:aff><country xml:lang="fr">Turquie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">27483984</idno><idno type="pmc">4971470</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4971470</idno><idno type="RBID">PMC:4971470</idno><idno type="doi">10.1038/srep30910</idno><date when="2016">2016</date><idno type="wicri:Area/Pmc/Corpus">000472</idno><idno type="wicri:Area/Pmc/Curation">000471</idno><idno type="wicri:Area/Pmc/Checkpoint">000114</idno><idno type="wicri:Area/Ncbi/Merge">002134</idno><idno type="wicri:Area/Ncbi/Curation">002134</idno><idno type="wicri:Area/Ncbi/Checkpoint">002134</idno><idno type="wicri:Area/Main/Merge">000139</idno><idno type="wicri:Area/Main/Curation">000139</idno><idno type="wicri:Area/Main/Exploration">000139</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Manipulating the Biosynthesis of Bioactive Compound Alkaloids for Next-Generation Metabolic Engineering in Opium Poppy Using CRISPR-Cas 9 Genome Editing Technology</title><author><name sortKey="Alagoz, Yagiz" sort="Alagoz, Yagiz" uniqKey="Alagoz Y" first="Yagiz" last="Alagoz">Yagiz Alagoz</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Biology, Faculty of Science, Çankırı Karatekin University</institution>, Çankırı18100,<country>Turkey</country></nlm:aff><country xml:lang="fr">Turquie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Hawkesbury Institute for the Environment, Western Sydney University</institution>, Richmond, New South Wales 2753,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Gurkok, Tugba" sort="Gurkok, Tugba" uniqKey="Gurkok T" first="Tugba" last="Gurkok">Tugba Gurkok</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Biology, Faculty of Science, Çankırı Karatekin University</institution>, Çankırı18100,<country>Turkey</country></nlm:aff><country xml:lang="fr">Turquie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Zhang, Baohong" sort="Zhang, Baohong" uniqKey="Zhang B" first="Baohong" last="Zhang">Baohong Zhang</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Department of Biology, East Carolina University</institution>, Greenville, NC 27858,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Unver, Turgay" sort="Unver, Turgay" uniqKey="Unver T" first="Turgay" last="Unver">Turgay Unver</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Department of Biology, Faculty of Science, Çankırı Karatekin University</institution>, Çankırı18100,<country>Turkey</country></nlm:aff><country xml:lang="fr">Turquie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Izmir International Biomedicine and Genome Institute (iBG-izmir), Dokuz Eylul University</institution>, Balcova 35340 Izmir,<country>Turkey</country></nlm:aff><country xml:lang="fr">Turquie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated9 (Cas9) endonuclease system is a powerful RNA-guided genome editing tool. CRISPR/Cas9 has been well studied in model plant species for targeted genome editing. However, few studies have been reported on plant species without whole genome sequence information. Currently, no study has been performed to manipulate metabolic pathways using CRISPR/Cas9. In this study, the type II CRISPR/SpCas9 system was used to knock out, via nonhomologous end-joining genome repair, the <italic>4</italic>′<italic>OMT2</italic> in opium poppy (<italic>Papaver somniferum</italic> L.), a gene which regulates the biosythesis of benzylisoquinoline alkaloids (BIAs). For sgRNA transcription, viral-based TRV and synthetic binary plasmids were designed and delivered into plant cells with a Cas9 encoding-synthetic vector by <italic>Agrobacterium</italic>-mediated transformation. InDels formed by CRISPR/Cas9 were detected by sequence analysis. Our results showed that the biosynthesis of BIAs (e.g. morphine, thebaine) was significantly reduced in the transgenic plants suggesting that <italic>4</italic>′<italic>OMT2</italic> was efficiently knocked-out by our CRISPR-Cas9 genome editing approach. In addition, a novel uncharacterized alkaloid was observed only in CRISPR/Cas9 edited plants. Thus, the applicabilitiy of the CRISPR/Cas9 system was demonstrated for the first time for medicinal aromatic plants by sgRNAs transcribed from both synthetic and viral vectors to regulate BIA metabolism and biosynthesis.</p></div></front><back><div1 type="bibliography"><listBibl><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><author><name sortKey="Li, X Q" uniqKey="Li X">X. Q. Li</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gaj, T" uniqKey="Gaj T">T. 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Ran</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sternberg, S H" uniqKey="Sternberg S">S. H. Sternberg</name></author><author><name sortKey="Redding, S" uniqKey="Redding S">S. Redding</name></author><author><name sortKey="Jinek, M" uniqKey="Jinek M">M. Jinek</name></author><author><name sortKey="Greene, E C" uniqKey="Greene E">E. C. Greene</name></author><author><name sortKey="Doudna, J A" uniqKey="Doudna J">J. A. Doudna</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Australie</li><li>Turquie</li><li>États-Unis</li></country></list><tree><country name="Turquie"><noRegion><name sortKey="Alagoz, Yagiz" sort="Alagoz, Yagiz" uniqKey="Alagoz Y" first="Yagiz" last="Alagoz">Yagiz Alagoz</name></noRegion><name sortKey="Gurkok, Tugba" sort="Gurkok, Tugba" uniqKey="Gurkok T" first="Tugba" last="Gurkok">Tugba Gurkok</name><name sortKey="Unver, Turgay" sort="Unver, Turgay" uniqKey="Unver T" first="Turgay" last="Unver">Turgay Unver</name><name sortKey="Unver, Turgay" sort="Unver, Turgay" uniqKey="Unver T" first="Turgay" last="Unver">Turgay Unver</name></country><country name="Australie"><noRegion><name sortKey="Alagoz, Yagiz" sort="Alagoz, Yagiz" uniqKey="Alagoz Y" first="Yagiz" last="Alagoz">Yagiz Alagoz</name></noRegion></country><country name="États-Unis"><noRegion><name sortKey="Zhang, Baohong" sort="Zhang, Baohong" uniqKey="Zhang B" first="Baohong" last="Zhang">Baohong Zhang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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