Journal Article Research Support, Non-U.S. Gov't Ppublish
PubMed: 38768032 DOI: 10.1016/j.xpro.2024.103072
Protocol to efficiently induce CRISPR-Kill-mediated cell ablation in Arabidopsis thaliana.
STAR Protoc 5(2):103072
Authors: Fabienne Gehrke, Sebastian Wolf, Holger Puchta
Affiliations
- Joseph Gottlieb Kölreuter Institute for Plant Sciences (JKIP) - Molecular Biology, Karlsruhe Institute of Technology (KIT), 76131 Karlsruhe, Germany.
- Centre for Plant Molecular Biology (ZMBP), Plant Biochemistry, Eberhard-Karls-University Tübingen, 72076 Tübingen, Germany.
- Joseph Gottlieb Kölreuter Institute for Plant Sciences (JKIP) - Molecular Biology, Karlsruhe Institute of Technology (KIT), 76131 Karlsruhe, Germany.
Abstract
The CRISPR-Kill system enables targeted cell ablation by inducing multiple double-strand breaks in evolutionarily conserved repetitive genomic regions. Here, we present a protocol for the application of the CRISPR-Kill system to analyze the systemic and cellular effects of targeted cell death in Arabidopsis. We describe steps for generating constitutive and inducible CRISPR-Kill lines, chemically inducing CRISPR-Cas9-mediated genome elimination, and monitoring of cell death in shoot and root apical meristems. This enables the investigation of a wide range of questions in developmental plant biology. For complete details on the use and execution of this protocol, please refer to Gehrke et al. 1.
MeSH
Cells, Clustered Regularly Interspaced Short Palindromic Repeats, Clinical Laboratory Techniques, Cell Death