The New phytologistOct 18, 2026

Adding uORFs in wheat increases tolerance to environmental stress without affecting growth

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Oct 19, 2026

Abstract

A total of 1199 actively translated upstream open reading frames were identified across the wheat genome.

Regulating gene expression in allohexaploid wheat can be achieved by disrupting or creating new upstream open reading frames through genome editing.
CRISPR/Cas9-mediated disruption of specific upstream open reading frames enhanced protein levels, which may improve drought, cold, and salt tolerance in plants.
Base editing techniques successfully created new upstream open reading frames that downregulated gene expression and potentially increased drought resilience.
The transgene-free uORF_TaGGP1-ACG and uORF_TaMADS26 mutants showed improved yields under drought without negative effects on growth in well-watered conditions.

Simplified

Developing climate-resilient crops is critical for ensuring global food security, yet agricultural productivity is frequently constrained by the trade-off between stress tolerance and growth. This challenge is particularly acute in allohexaploid wheat, a staple crop susceptible to abiotic stresses, underscoring the demand for precise gene regulation. Here, we established a simple, efficient, and transgene-free method for regulating gene expression in allohexaploid wheat by disrupting or creating de novo upstream open reading frames (uORFs) via genome editing. Using ribosome profiling, we identified 1199 actively translated uORFs across the wheat genome and developed a dual-luciferase screening pipeline to pinpoint potent translational repressors of positive regulators of abiotic stress. CRISPR/Cas9-mediated disruption of the uORFs in TaCIPK14 and TaGGP1 elevated downstream protein accumulation, thereby enhancing drought, cold, and salt tolerance of plants. Furthermore, base editing was employed to generate de novo uORFs in TaMADS26, effectively downregulating gene expression and improving drought resilience. Notably, both the transgene-free uorf_TaGGP1-ACG and uORF_TaMADS26 mutants conferred pronounced yield advantages under drought conditions without incurring growth penalties under well-watered conditions. This study provides a predictable and precise way for modulating gene expression, enabling the coordinated improvement of stress resilience and agronomic performance in wheat.

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Adding uORFs in wheat increases tolerance to environmental stress without affecting growth

Abstract

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