Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation |
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Authors: | Zhi-Ping Wang Hui-Li Xing Li Dong Hai-Yan Zhang Chun-Yan Han Xue-Chen Wang Qi-Jun Chen |
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Affiliation: | State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193 China |
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Abstract: | Arabidopsis mutants produced by constitutive overexpression of the CRISPR/Cas9 genome editing system are usually mosaics in the T1 generation. In this study, we used egg cell-specific promoters to drive the expression of Cas9 and obtained non-mosaic T1 mutants for multiple target genes with high efficiency. Comparisons of 12 combinations of eight promoters and two terminators found that the efficiency of the egg cell-specific promoter-controlled CRISPR/Cas9 system depended on the presence of a suitable terminator, and the composite promoter generated by fusing two egg cell-specific promoters resulted in much higher efficiency of mutation in the T1 generation compared with the single promoters.Electronic supplementary materialThe online version of this article (doi:10.1186/s13059-015-0715-0) contains supplementary material, which is available to authorized users. |
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