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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.accessioned2017-08-28T10:28:00Z
dc.date.available2017-08-28T10:28:00Z
dc.date.issued2017-08-24
dc.identifier.citationButt H, Eid A, Ali Z, Atia MAM, Mokhtar MM, et al. (2017) Efficient CRISPR/Cas9-Mediated Genome Editing Using a Chimeric Single-Guide RNA Molecule. Frontiers in Plant Science 8. Available: http://dx.doi.org/10.3389/fpls.2017.01441.
dc.identifier.issn1664-462X
dc.identifier.pmid28883826
dc.identifier.doi10.3389/fpls.2017.01441
dc.identifier.urihttp://hdl.handle.net/10754/625405
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.description.sponsorshipThis study was supported by King Abdullah University of Science and Technology.
dc.publisherFrontiers Media SA
dc.relation.urlhttp://journal.frontiersin.org/article/10.3389/fpls.2017.01441/full
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.urihttp://creativecommons.org/licenses/by/4.0/
dc.titleEfficient CRISPR/Cas9-Mediated Genome Editing Using a Chimeric Single-Guide RNA Molecule
dc.typeArticle
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Division
dc.contributor.departmentBioscience Program
dc.contributor.departmentDesert Agriculture Initiative
dc.contributor.departmentLaboratory for Genome Engineering
dc.contributor.departmentPlant Science
dc.contributor.departmentPlant Science Program
dc.identifier.journalFrontiers in Plant Science
dc.eprint.versionPublisher's Version/PDF
dc.contributor.institutionMolecular Genetics and Genome Mapping Laboratory, Agricultural Genetic Engineering Research Institute, Agricultural Research Center, Giza, Egypt
dc.contributor.institutionDepartment of Bioengineering, Rice University, Houston, TX, United States
kaust.personButt, Haroon
kaust.personEid, Ayman
kaust.personAli, Zahir
kaust.personHassan, Norhan
kaust.personMahfouz, Magdy M.
refterms.dateFOA2018-06-13T19:04:32Z


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This 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.
Except where otherwise noted, this item's license is described as This 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.