Efficient CRISPR/Cas9-Mediated Genome Editing Using a Chimeric Single-Guide RNA Molecule
Atia, Mohamed A. M.
Mokhtar, Morad M.
Lee, Ciaran M.
Mahfouz, Magdy M.
KAUST DepartmentBiological and Environmental Sciences and Engineering (BESE) Division
Plant Science Program
Desert Agriculture Initiative
Laboratory for Genome Engineering, Division of Biological Sciences, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia
Permanent link to this recordhttp://hdl.handle.net/10754/625405
MetadataShow full item record
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.
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.
SponsorsThis study was supported by King Abdullah University of Science and Technology.
PublisherFrontiers Media SA
JournalFrontiers in Plant Science
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.
- Efficient introduction of specific homozygous and heterozygous mutations using CRISPR/Cas9.
- Authors: Paquet D, Kwart D, Chen A, Sproul A, Jacob S, Teo S, Olsen KM, Gregg A, Noggle S, Tessier-Lavigne M
- Issue date: 2016 May 5
- Precision genome editing in the CRISPR era.
- Authors: Salsman J, Dellaire G
- Issue date: 2017 Apr
- CRISPR-Cas9-Mediated Genome Editing in Leishmania donovani.
- Authors: Zhang WW, Matlashewski G
- Issue date: 2015 Jul 21
- Highly specific and efficient CRISPR/Cas9-catalyzed homology-directed repair in Drosophila.
- Authors: Gratz SJ, Ukken FP, Rubinstein CD, Thiede G, Donohue LK, Cummings AM, O'Connor-Giles KM
- Issue date: 2014 Apr
- CRISPR-Cas9 Genome Editing in Drosophila.
- Authors: Gratz SJ, Rubinstein CD, Harrison MM, Wildonger J, O'Connor-Giles KM
- Issue date: 2015 Jul 1