A versatile CRISPR-based system for lineage tracing in living plants

Area of Science:

• Developmental Biology
• Plant Science
• Genetics

Background:

• Understanding cell lineage diversification is crucial in multicellular organism development.
• Existing lineage tracing methods have limitations in plants.
• CRISPR technology offers potential for advanced genetic manipulation.

Purpose of the Study:

• To develop a novel, single-reporter CRISPR-based system for plant cell lineage tracing.
• To enable precise documentation of cell lineages in living plants.
• To create a versatile system applicable across different plant species.

Main Methods:

• Exploiting CRISPR-Cas9 mutagenic activity to correct a frameshift mutation in a reporter gene.
• Restoring nuclear fluorescent protein expression to label initial cells and their progeny.
• Utilizing tissue-specific and inducible promoters for spatial and temporal control of Cas9 activity.
• Demonstrating proof of principle in two model plant systems.

Main Results:

• Successfully established a CRISPR-induced lineage tracing system in plants.
• The system labels initial cells and their descendants with a strong fluorescent signal.
• Cas9-induced mutations did not affect other plant phenotypes.
• Demonstrated functionality in two distinct model plants.

Conclusions:

• The developed CRISPR system provides an effective and non-disruptive method for plant cell lineage tracing.
• The system's conserved components and adaptable cloning facilitate broad application in plant developmental biology.
• This technique advances the study of cell lineage contributions to mature plant organisms.