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Protocol to efficiently induce CRISPR-Kill-mediated cell ablation in Arabidopsis thaliana
Fabienne Gehrke1, Sebastian Wolf2, Holger Puchta1
1Joseph Gottlieb Kölreuter Institute for Plant Sciences (JKIP) - Molecular Biology, Karlsruhe Institute of Technology (KIT), 76131 Karlsruhe, Germany.
STAR Protocols
|May 20, 2024
Summary
The CRISPR-Kill system precisely eliminates plant cells by targeting repetitive DNA. This protocol details its use in Arabidopsis for studying cell death effects in plant development.
Area of Science:
- Plant Biology
- Genetics
- Molecular Biology
Background:
- Targeted cell ablation is crucial for understanding developmental processes.
- CRISPR-Cas9 technology offers precise genomic manipulation.
Purpose of the Study:
- To present a detailed protocol for applying the CRISPR-Kill system in Arabidopsis.
- To enable the analysis of systemic and cellular effects of targeted cell death.
Main Methods:
- Generating constitutive and inducible CRISPR-Kill lines.
- Chemically inducing CRISPR-Cas9-mediated genome elimination.
- Monitoring cell death in shoot and root apical meristems.
Main Results:
- The protocol facilitates the study of targeted cell death in plants.
- CRISPR-Kill system effectively induces double-strand breaks in repetitive genomic regions.
- Allows for investigation of developmental plant biology questions.
Conclusions:
- The CRISPR-Kill system is a valuable tool for plant developmental research.
- This protocol provides a framework for analyzing cell death mechanisms.
- Enables broad applications in studying plant growth and development.
