Botanical Institute - Molecular Biology and Biochemistry of Plants, Karlsruhe Institute of Technology, Fritz-Haber-Weg 4, 76131, Karlsruhe, Germany.
Nat Commun. 2022 Mar 21;13(1):1502. doi: 10.1038/s41467-022-29130-w.
CRISPR/Cas has been mainly used for mutagenesis through the induction of double strand breaks (DSBs) within unique protein-coding genes. Using the SaCas9 nuclease to induce multiple DSBs in functional repetitive DNA of Arabidopsis thaliana, we can now show that cell death can be induced in a controlled way. This approach, named CRISPR-Kill, can be used as tool for tissue engineering. By simply exchanging the constitutive promoter of SaCas9 with cell type-specific promoters, it is possible to block organogenesis in Arabidopsis. By AP1-specific expression of CRISPR-Kill, we are able to restore the apetala1 phenotype and to specifically eliminate petals. In addition, by expressing CRISPR-Kill in root-specific pericycle cells, we are able to dramatically reduce the number and the length of lateral roots. In the future, the application of CRISPR-Kill may not only help to control development but could also be used to change the biochemical properties of plants.
CRISPR/Cas 主要通过在独特的蛋白质编码基因内诱导双链断裂 (DSB) 来进行突变。使用 SaCas9 核酸酶在拟南芥的功能重复 DNA 中诱导多个 DSB，我们现在可以证明可以以可控的方式诱导细胞死亡。这种方法称为 CRISPR-Kill，可用作组织工程工具。通过简单地将 SaCas9 的组成型启动子与细胞类型特异性启动子交换，就可以阻止拟南芥的器官发生。通过 AP1 特异性表达 CRISPR-Kill，我们能够恢复拟南芥的无花瓣表型并特异性地消除花瓣。此外，通过在根特异性周细胞中表达 CRISPR-Kill，我们能够显著减少侧根的数量和长度。将来，CRISPR-Kill 的应用不仅有助于控制发育，还可以用于改变植物的生化特性。
