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Chemical mutagenesis continues to be a widely used method for introducing genetic diversity in plant breeding programs due to its ability to generate point mutations without triggering GMO regulatory classification. While such mutations in coding regions can improve specific traits, they may simultaneously reduce the protein’s endogenous function, potentially impacting overall plant fitness. In such cases, compensatory mechanisms may be required to restore function or maintain viability.
One promising approach is to increase the expression level of the mutated gene by altering its promoter region. Even single-nucleotide changes in promoter sequences can significantly affect transcriptional activity, offering a route to fine-tune gene expression in a non-transgenic manner.
Understanding the relationship between specific promoter variants and expression outcomes is therefore key. Innovative approaches that enable the functional characterization of single-nucleotide mutations in promoter regions would improve understanding of how natural or induced regulatory variation impacts gene expression, and support the identification of mutant plant lines carrying favorable promoter variants that enhance the expression of target genes.
We are seeking to collaborate with an R&D partner to investigate how single-nucleotide changes in promoter regions affect the expression levels of their associated genes. This may involve leveraging existing sequence diversity from mutant populations or germplasm panels, or creating novel variation through synthetic promoter libraries. Of particular interest are methodologies that allow functional characterization of these regulatory variants, linking specific promoter mutations to measurable expression outcomes.
- Study designs to assess single nucleotide polymorphisms (SNP) impact in promoter regions (e.g., expression-linked assays)
- Reporter-based systems for promoter variant analysis (e.g., β-glucuronidase, luciferase, fluorescent reporters)
- High-throughput assays linking promoter variants to expression outcomes (e.g., transient assays, dual-luciferase systems)
- High-throughput expression quantification tools (e.g., qPCR, RNA-seq, imaging assays)
- In silico analysis of mutant population regarding the occurrence and frequency of SNPs causing changes in expression level from a real live example
- Leverages promoter sequence diversity derived from mutant populations, germplasm panels, or synthetic promoter libraries
- Assesses the likelihood of successfully implementing the above screening approach in plant using a mutagenesis approach
- Compatible with high-throughput or multiplexed analysis
- Preliminary data showing clear links between promoter sequence and expression
- Screening-only platforms that do not assess or interpret promoter variant function
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