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

• Published in: Plants (Basel) Vol. 12; no. 12; p. 2365
• Main Authors: Li, Meng, Niu, Xufang, Li, Shuang, Fu, Shasha, Li, Qianfang, Xu, Meizhi, Wang, Chunhua, Wu, Shuang
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 19-06-2023; MDPI
• Subjects: Cell division; cell-type-specific gene knock-out; CRISPR; CRISPR/Cas9; Functionals; Genes; genes function; Genomics; Gibberellic acid; Homeobox; Lethality; Mutagenesis; Mutation; Phenotypes; Proteins; root; Signal transduction; Stem cells; cell-type-specific gene knock-out; genes function; CRISPR/Cas9; root
• ISSN: 2223-7747; 2223-7747
• DOI: 10.3390/plants12122365

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
(
),
(
), and
(
) 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
(
) and
(
) 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.