Abstract

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.

MeSH

Biological Science Disciplines, Clustered Regularly Interspaced Short Palindromic Repeats, Tandem Repeat Sequences, Cells, Genes, Promoter Regions, Genetic

Cited by

• Protocol to efficiently induce CRISPR-Kill-mediated cell ablation in Arabidopsis thaliana. STAR Protoc, 2024
• Centromeres: From chromosome biology to biotechnology applications and synthetic genomes in plants. Plant Biotechnol J, 2022
• Low dose ribosomal DNA P-loop mutation affects development and enforces autophagy in Arabidopsis. RNA Biol, 2024
• Conditional knockdown of gene expression in plants via 3' UTR editing. Plant Commun, 2025
• Centromere diversity and its evolutionary impacts on plant karyotypes and plant reproduction. New Phytol, 2025
• Combining Multiplexed CRISPR/Cas9-Nickase and PARP Inhibitors Efficiently and Precisely Targets Cancer Cells. Cancer Res, 2025