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Monitoring the stability of transgene expression in lettuce using the RUBY reporter.
Beth A Rowan1,2,3, Megan Reeves4,5, Claire Hays4,5,6,7
1The Genome Center, University of California, Davis, Davis, CA, USA. beth.rowan@oregonstate.edu.
Plant Cell Reports
|November 25, 2025
Summary
The lettuce ubiquitin promoter and terminator offer strong, stable transgene expression across generations, outperforming the Cauliflower Mosaic Virus 35S promoter in lettuce. This finding provides valuable tools for developing stable transgenic lettuce.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Genetics
Background:
- Stable transgene expression in lettuce remains a challenge, with common promoters like Cauliflower Mosaic Virus 35S showing instability.
- Limited research exists on various promoter/terminator combinations for stable transgene expression in lettuce.
Purpose of the Study:
- To evaluate and compare the efficacy of different transgene expression constructs in lettuce.
- To identify robust promoter and terminator combinations for stable and strong transgene expression in lettuce.
Main Methods:
- Utilized the non-invasive RUBY reporter system for monitoring transgene expression.
- Assessed three distinct expression constructs (LsUBI, AtUBI/tRBCS, 35S/tHSP) in two lettuce varieties.
- Tracked transgene expression during regeneration, primary transgenic development, and two subsequent generations.
Main Results:
- The LsUBI promoter/terminator combination demonstrated strong, uniform, and stable expression across all stages and generations.
- The AtUBI promoter/tRBCS combination showed slightly lower expression with intermediate silencing.
- The 35S promoter/tHSP combination exhibited initial strong expression but frequent silencing.
Conclusions:
- The LsUBI promoter/terminator is highly effective for strong, uniform, and stable transgene expression in lettuce, minimizing silencing.
- The AtUBI promoter/tRBCS combination offers a viable alternative for stable transgene expression in lettuce, particularly when intermediate expression levels are desired.
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