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

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

Bibliography:

Ayman Eid, Zahir Ali, Magdy M. Mahfouz. High efficiency of targeted mutagenesis in arabidopsis via meiotic promoter-driven expression of Cas9 endonuclease. Plant Cell Reports. DOI: 10.1007/s00299-016-2000-4

Authors:

Ayman Eid, Zahir Ali, Magdy M. Mahfouz

Keywords:

CRISPR/Cas9,Genome engineering, Genome editing, Meiotic promoters, Targeted mutagenesis, Arabidopsis

Year:

2016

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.

ISSN:

0721-7714