Efficient CRISPR/Cas9-Mediated Genome Editing Using a Chimeric Single-Guide RNA Molecule

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dc.citation.articleNumber1441en_US
dc.citation.journalTitleFrontiers in Plant Scienceen_US
dc.citation.volumeNumber8en_US
dc.contributor.authorButt, Haroonen_US
dc.contributor.authorEid, Aymanen_US
dc.contributor.authorAli, Zahiren_US
dc.contributor.authorAtia, Mohamed A.M.en_US
dc.contributor.authorMokhtar, Morad M.en_US
dc.contributor.authorHassan, Norhanen_US
dc.contributor.authorLee, Ciaran M.en_US
dc.contributor.authorBao, Gangen_US
dc.contributor.authorMahfouz, Magdy M.en_US
dc.contributor.orgBioengineeringen_US
dc.date.accessioned2017-11-15T14:12:53Zen_US
dc.date.available2017-11-15T14:12:53Zen_US
dc.date.issued2017en_US
dc.description.abstractThe CRISPR/Cas9 system has been applied in diverse eukaryotic organisms for targeted mutagenesis. However, targeted gene editing is inefficient and requires the simultaneous delivery of a DNA template for homology-directed repair (HDR). Here, we used CRISPR/Cas9 to generate targeted double-strand breaks and to deliver an RNA repair template for HDR in rice (Oryza sativa). We used chimeric single-guide RNA (cgRNA) molecules carrying both sequences for target site specificity (to generate the double-strand breaks) and repair template sequences (to direct HDR), flanked by regions of homology to the target. Gene editing was more efficient in rice protoplasts using repair templates complementary to the non-target DNA strand, rather than the target strand. We applied this cgRNA repair method to generate herbicide resistance in rice, which showed that this cgRNA repair method can be used for targeted gene editing in plants. Our findings will facilitate applications in functional genomics and targeted improvement of crop traits.en_US
dc.identifier.citationButt, Haroon, Eid, Ayman, Ali, Zahir, et al.. "Efficient CRISPR/Cas9-Mediated Genome Editing Using a Chimeric Single-Guide RNA Molecule." <i>Frontiers in Plant Science,</i> 8, (2017) Frontiers Media S.A.: https://doi.org/10.3389/fpls.2017.01441.en_US
dc.identifier.digitalCRISPR_Cas9-Mediated_Genome_Editingen_US
dc.identifier.doihttps://doi.org/10.3389/fpls.2017.01441en_US
dc.identifier.urihttps://hdl.handle.net/1911/98824en_US
dc.language.isoengen_US
dc.publisherFrontiers Media S.A.en_US
dc.rightsThis is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.en_US
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en_US
dc.subject.keywordCRISPR/Cas9en_US
dc.subject.keywordHDRen_US
dc.subject.keywordRNA-templated repairen_US
dc.subject.keywordgene editingen_US
dc.subject.keywordgenome engineeringen_US
dc.titleEfficient CRISPR/Cas9-Mediated Genome Editing Using a Chimeric Single-Guide RNA Moleculeen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpublisher versionen_US
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