Optimization of a hypercompact Fanzor2 system for improved genome editing performance in plants

Area of Science:

• Plant biotechnology
• Molecular biology
• Genome editing

Background:

• Native Fanzor2 nucleases have limited editing efficiency in plant systems.
• Targeting recalcitrant genomic sites remains a challenge for current gene editing tools.

Purpose of the Study:

• To engineer an optimized Fanzor2 system for robust plant genome editing.
• To enable precise cytosine base editing in crops.
• To support versatile crop genome modification strategies.

Main Methods:

• ωRNA optimization of Fanzor2 nucleases.
• Structure-based protein mutagenesis.
• Evaluation of editing activity at recalcitrant genomic sites.
• Cytosine base editing assays.
• Transformation and viral delivery methods.

Main Results:

• Engineered Fanzor2 system demonstrated significantly improved editing activity in plants.
• Achieved robust editing at previously recalcitrant genomic loci.
• Successfully implemented cytosine base editing using the optimized system.
• Demonstrated versatility through both transformation and viral delivery.

Conclusions:

• The optimized Fanzor2 system overcomes limitations of native nucleases for plant genome editing.
• This technology provides a powerful tool for versatile crop genome modification, including precise base editing.
• The system supports multiple delivery methods, enhancing its applicability in crop improvement programs.