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

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dc.citation.articleNumber1441
dc.citation.journalTitleFrontiers in Plant Science
dc.citation.volumeNumber8
dc.contributor.authorButt, Haroon
dc.contributor.authorEid, Ayman
dc.contributor.authorAli, Zahir
dc.contributor.authorAtia, Mohamed A.M.
dc.contributor.authorMokhtar, Morad M.
dc.contributor.authorHassan, Norhan
dc.contributor.authorLee, Ciaran M.
dc.contributor.authorBao, Gang
dc.contributor.authorMahfouz, Magdy M.
dc.date.accessioned2017-11-15T14:12:53Z
dc.date.available2017-11-15T14:12:53Z
dc.date.issued2017
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.
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.
dc.identifier.digitalCRISPR_Cas9-Mediated_Genome_Editing
dc.identifier.doihttps://doi.org/10.3389/fpls.2017.01441
dc.identifier.urihttps://hdl.handle.net/1911/98824
dc.language.isoeng
dc.publisherFrontiers Media S.A.
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.
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subject.keywordCRISPR/Cas9
dc.subject.keywordHDR
dc.subject.keywordRNA-templated repair
dc.subject.keywordgene editing
dc.subject.keywordgenome engineering
dc.titleEfficient CRISPR/Cas9-Mediated Genome Editing Using a Chimeric Single-Guide RNA Molecule
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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