Engineering disease-resistant plants with alternative translation efficiency by switching uORF types through CRISPR

Area of Science:

• Agricultural Science
• Molecular Biology
• Genetics

Background:

• Engineering disease-resistant plants is crucial for food security.
• Controlling gene expression is key to plant disease resistance.
• Upstream open reading frames (uORFs) regulate gene translation.

Purpose of the Study:

• To optimize translational control of disease-related genes in rice using CRISPR-edited uORF variants.
• To develop a method for pinpointing genes with optimal expression levels for disease resistance.
• To enhance crop resilience against diseases.

Main Methods:

• Screening of CRISPR-edited uORF variants in rice.
• Modulating ribosome accessibility to downstream genes by switching uORF types.
• Utilizing firefly luciferase as a reporter system.
• Generating alternative translation efficiency uORF variants (CRISPR-aTrE-uORF).

Main Results:

• Enhanced translational suppression of glutamine synthetase 2 provided broad-spectrum disease resistance in rice.
• Minimal fitness costs were observed with the engineered resistance.
• The strategy demonstrated the potential of modifying uORFs to control gene expression.

Conclusions:

• Translational control via uORF engineering is a viable strategy for developing disease-resistant crops.
• This approach can accelerate the development of crops to meet food security demands.
• Precision control of gene translation offers a novel pathway for agricultural innovation.