Chimeric Regulatory Sequences for Plant Gene Expression
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Solution Overview
Problem
Current plant genetic engineering techniques face challenges in achieving reproducible and optimal control of gene expression in plants, particularly when introducing multiple genes, as they often result in gene silencing and variable expression patterns due to factors like insertion site, regulatory sequence strength, and codon usage.
Innovation Solution
The use of chimeric regulatory sequences comprising a monocot promoter and a dicot intron enhances and broadens the expression pattern of transgenes, allowing for improved modulation of gene expression by incorporating a dicot intron derived from a dicot species into a monocot promoter, which is fused with a structural gene within a DNA plant expression construct.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple genes are introduced into crop plants using conventional genetic engineering techniques, then the plants acquire multiple desired traits, but gene silencing and variable expression patterns occur due to factors like insertion site, regulatory sequence strength, and codon usage
Solution Approach 1:
The patent employs a composite regulatory sequence comprising a promoter from one plant species and an intron from a different plant species (cross-kingdom intron-promoter combination). This composite structure integrates elements from different biological sources to create a regulatory module that overcomes the limitations of conventional single-source regulatory sequences, thereby reducing gene silencing and improving expression reproducibility across multiple introduced genes
Solution Approach 2:
The patent systematically varies key parameters of the regulatory sequence, specifically using introns from different species (dicot vs. monocot) and promoters from different species to optimize gene expression. By changing the biological origin and structural parameters of the regulatory elements, the patent achieves more consistent and predictable expression patterns for multiple introduced genes
2Ease of operation
If a single regulatory sequence is used to control transgene expression, then the construct is simple, but the expression pattern is limited and cannot be optimized for different tissues or conditions
Solution Approach 1:
The patent creates a composite regulatory sequence by combining a promoter element with an intron element from different biological kingdoms. This composite structure maintains relative simplicity in the overall construct design while achieving enhanced and broadened expression patterns across different plant tissues and conditions, thus resolving the contradiction between simplicity and versatility
Data Source
AI summary
The present invention relates to a method of using a dicot intron or elements thereof to enhance transgene expression in plants. The present invention also provides constructs, transgenic plants and seeds containing the polynucleotide useful for expressing transgene in plants.