Synthetic Intergenic Sequence Regions for Stable Transgene Stacking
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
In plant biotechnology, the interaction of expression elements from one transgene cassette can alter the expression profile of another cassette in a vector stack, affecting the intended phenotype, particularly in seed-specific expression patterns.
Innovation Solution
The use of novel synthetic Intergenic Sequence Regions (ISRs) inserted between transgene cassettes in a vector stack to minimize interaction, preserving the intended expression pattern and level of each cassette.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiple transgene cassettes are cloned into a single vector stack to introduce several agronomic traits, then breeding efficiency is improved and cost is reduced, but expression elements from one cassette can alter the expression profile of another cassette
Solution Approach 1:
The patent introduces Intergenic Sequence Regions (ISRs) as intermediary DNA sequences placed between adjacent transgene cassettes in a vector stack. These ISRs act as buffers that reduce or eliminate the interaction between expression elements of neighboring cassettes, thereby preventing unwanted alterations in expression profiles while maintaining the benefits of multi-trait stacking.
2Adaptability or versatility
If a seed-specific expression cassette is placed in a vector stack with other cassettes, then seed-specific expression is achieved, but interaction with neighboring expression elements can alter the seed-specific expression pattern to a more constitutive pattern
Solution Approach 1:
ISRs are positioned between the seed-specific expression cassette and adjacent cassettes to serve as protective buffers. These intermediary sequences reduce the influence of neighboring expression elements on the seed-specific cassette, thereby preserving the fidelity of the intended seed-specific expression pattern and preventing unwanted constitutive expression.
Solution Approach 2:
The patent applies different ISR sequences strategically between specific pairs of cassettes based on their expression patterns and potential interactions. This localized approach allows optimization of expression fidelity for each specific cassette configuration while maintaining overall system functionality.
3Device complexity
If no intergenic sequence is placed between transgene cassettes, then vector complexity is reduced, but expression elements from one cassette interact with and alter the expression of adjacent cassettes
Solution Approach 1:
The patent introduces ISRs as minimal intermediary sequences that provide the necessary buffering function with relatively small additions to vector complexity. These ISRs are specifically designed to reduce interaction between cassettes while maintaining a manageable increase in overall vector structure.
Data Source
AI summary
The invention provides recombinant DNA molecules comprising novel synthetic Intergenic Sequence Regions for use in plants to reduce the interaction of a first transgene expression cassette on a second transgene cassette when inserted between the first transgene cassette and second transgene cassette. The invention also provides transgenic plants, plant cells, plant parts, and seeds comprising the novel synthetic Intergenic Sequence Regions. The invention also provides methods to reduce the interaction between transgene expression cassettes using the novel synthetic Intergenic Sequence Regions.