CRISPR/Cas9 Based Cell-Type Specific Gene Knock-Out in Arabidopsis Roots

被引:2
作者
Li, Meng
Niu, Xufang
Li, Shuang
Fu, Shasha
Li, Qianfang
Xu, Meizhi
Wang, Chunhua
Wu, Shuang [1 ]
机构
[1] Fujian Agr & Forestry Univ, Coll Life Sci, Fuzhou 350002, Peoples R China
来源
PLANTS-BASEL | 2023年 / 12卷 / 12期
关键词
CRISPR; Cas9; genes function; cell-type-specific gene knock-out; root; STRAND BREAK REPAIR; TISSUE; GIBBERELLIN; ORGANIZATION; MUTAGENESIS; SCARECROW; SYSTEM; TSKO;
D O I
10.3390/plants12122365
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
CRISPR/Cas9 (hereafter Cas9)-mediated gene knockout is one of the most important tools for studying gene function. However, many genes in plants play distinct roles in different cell types. Engineering the currently used Cas9 system to achieve cell-type-specific knockout of functional genes is useful for addressing the cell-specific functions of genes. Here we employed the cell-specific promoters of the WUSCHEL RELATED HOMEOBOX 5 (WOX5), CYCLIND6;1 (CYCD6;1), and ENDODERMIS7 (EN7) genes to drive the Cas9 element, allowing tissue-specific targeting of the genes of interest. We designed the reporters to verify the tissue-specific gene knockout in vivo. Our observation of the developmental phenotypes provides strong evidence for the involvement of SCARECROW (SCR) and GIBBERELLIC ACID INSENSITIVE (GAI) in the development of quiescent center (QC) and endodermal cells. This system overcomes the limitations of traditional plant mutagenesis techniques, which often result in embryonic lethality or pleiotropic phenotypes. By allowing cell-type-specific manipulation, this system has great potential to help us better understand the spatiotemporal functions of genes during plant development.
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页数:11
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