title: Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation.
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.
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China. wangzp6119@163.com.
authors.forename: Zhi-Ping
authors.initials: ZP
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China. xinghuili1989@163.com.
authors.forename: Hui-Li
authors.initials: HL
authors.lastname: Xing
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China. li.dong127@gmail.com.
authors.forename: Li
authors.initials: L
authors.lastname: Dong
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China. hzzhanghaiyan@163.com.
authors.forename: Hai-Yan
authors.initials: HY
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China. qy15652815165@163.com.
authors.forename: Chun-Yan
authors.initials: CY
authors.lastname: Han
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China. xuechen.wang@gmail.com.
authors.forename: Xue-Chen
authors.initials: XC
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China. qjchen@cau.edu.cn.
authors.forename: Qi-Jun
authors.initials: QJ
authors.lastname: Chen
mesh_terms: D017360:Arabidopsis / Q000196:embryology / Q000235:genetics*; D064130:CRISPR-Associated Proteins / Q000235:genetics / Q000378:metabolism; D064113:CRISPR-Cas Systems; D017343:Genes, Plant; D055993:Germ Cells, Plant / Q000378:metabolism; D006720:Homozygote; D009154:Mutation; D010641:Phenotype; D011401:Promoter Regions, Genetic; D013728:Terminator Regions, Genetic
chemical_list: D064130:CRISPR-Associated Proteins