High efficiency of targeted mutagenesis in arabidopsis via meiotic promoter-driven expression of Cas9 endonuclease

Handle URI:
http://hdl.handle.net/10754/621397
Title:
High efficiency of targeted mutagenesis in arabidopsis via meiotic promoter-driven expression of Cas9 endonuclease
Authors:
Eid, Ayman; Ali, Zahir ( 0000-0002-7814-8908 ) ; Mahfouz, Magdy M. ( 0000-0002-0616-6365 )
Abstract:
Key message: The use of a meiosis I-specific promoter increased the efficiency of targeted mutagenesis and will facilitate the manipulation of homologous recombination. Abstract: The CRISPR/Cas9 system has been harnessed for targeted engineering of eukaryotic genomes, including plants; however, CRISPR/Cas9 efficiency varies considerably in different plant tissues and species. In Arabidopsis, the generation of homozygous or bi-allelic mutants in the first (T1) generation is inefficient. Here, we used specific promoters to drive the expression of Cas9 during meiosis to maximize the efficiency of recovering heritable mutants in T1 plants. Our data reveal that the use of a promoter active in meiosis I resulted in high-efficiency (28 %) recovery of targeted mutants in the T1 generation. Moreover, this method enabled efficient simultaneous targeting of three genes for mutagenesis. Taken together, our results show that the use of meiosis-specific promoters will improve methods for functional genomic analysis and studying the molecular underpinnings of homologous recombination. © 2016, Springer-Verlag Berlin Heidelberg.
KAUST Department:
Laboratory for Genome Engineering, Division of Environmental and Biological Sciences and Engineering, King Abdullah University of Science and Technology, 4700, Thuwal, Saudi Arabia
Citation:
Eid A, Ali Z, Mahfouz MM (2016) High efficiency of targeted mutagenesis in arabidopsis via meiotic promoter-driven expression of Cas9 endonuclease. Plant Cell Reports 35: 1555–1558. Available: http://dx.doi.org/10.1007/s00299-016-2000-4.
Publisher:
Springer Nature
Journal:
Plant Cell Reports
Issue Date:
28-May-2016
DOI:
10.1007/s00299-016-2000-4
Type:
Article
ISSN:
0721-7714; 1432-203X
Sponsors:
We thank Qi-Jun Chen, State Key Laboratory of Plant Physiology and Biochemistry China Agricultural University, for providing the pHEE2A-TRI vector backbone. We thank members of the Laboratory for Genome Engineering at King Abdullah University of Science and Technology for helpful discussions and comments. The study was supported by the King Abdullah University of Science and Technology.
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorEid, Aymanen
dc.contributor.authorAli, Zahiren
dc.contributor.authorMahfouz, Magdy M.en
dc.date.accessioned2016-11-03T08:28:22Z-
dc.date.available2016-11-03T08:28:22Z-
dc.date.issued2016-05-28en
dc.identifier.citationEid A, Ali Z, Mahfouz MM (2016) High efficiency of targeted mutagenesis in arabidopsis via meiotic promoter-driven expression of Cas9 endonuclease. Plant Cell Reports 35: 1555–1558. Available: http://dx.doi.org/10.1007/s00299-016-2000-4.en
dc.identifier.issn0721-7714en
dc.identifier.issn1432-203Xen
dc.identifier.doi10.1007/s00299-016-2000-4en
dc.identifier.urihttp://hdl.handle.net/10754/621397-
dc.description.abstractKey message: The use of a meiosis I-specific promoter increased the efficiency of targeted mutagenesis and will facilitate the manipulation of homologous recombination. Abstract: The CRISPR/Cas9 system has been harnessed for targeted engineering of eukaryotic genomes, including plants; however, CRISPR/Cas9 efficiency varies considerably in different plant tissues and species. In Arabidopsis, the generation of homozygous or bi-allelic mutants in the first (T1) generation is inefficient. Here, we used specific promoters to drive the expression of Cas9 during meiosis to maximize the efficiency of recovering heritable mutants in T1 plants. Our data reveal that the use of a promoter active in meiosis I resulted in high-efficiency (28 %) recovery of targeted mutants in the T1 generation. Moreover, this method enabled efficient simultaneous targeting of three genes for mutagenesis. Taken together, our results show that the use of meiosis-specific promoters will improve methods for functional genomic analysis and studying the molecular underpinnings of homologous recombination. © 2016, Springer-Verlag Berlin Heidelberg.en
dc.description.sponsorshipWe thank Qi-Jun Chen, State Key Laboratory of Plant Physiology and Biochemistry China Agricultural University, for providing the pHEE2A-TRI vector backbone. We thank members of the Laboratory for Genome Engineering at King Abdullah University of Science and Technology for helpful discussions and comments. The study was supported by the King Abdullah University of Science and Technology.en
dc.publisherSpringer Natureen
dc.subjectArabidopsisen
dc.subjectCRISPR/Cas9en
dc.subjectGenome editingen
dc.subjectGenome engineeringen
dc.subjectMeiotic promotersen
dc.subjectTargeted mutagenesisen
dc.titleHigh efficiency of targeted mutagenesis in arabidopsis via meiotic promoter-driven expression of Cas9 endonucleaseen
dc.typeArticleen
dc.contributor.departmentLaboratory for Genome Engineering, Division of Environmental and Biological Sciences and Engineering, King Abdullah University of Science and Technology, 4700, Thuwal, Saudi Arabiaen
dc.identifier.journalPlant Cell Reportsen
kaust.authorEid, Aymanen
kaust.authorAli, Zahiren
kaust.authorMahfouz, Magdy M.en
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