Plant Terminator Sequences for Stable Transgene Expression
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Solution Overview
Problem
Current plant genetic engineering techniques face challenges in modulating gene expression in transgenic plants due to repetitive sequences, which can lead to undesirable effects on transgene expression and stability, necessitating the discovery of novel regulatory elements like terminator sequences for precise control of heterologous nucleic acid expression.
Innovation Solution
The use of novel terminator sequences, such as those identified in Sorghum bicolor and Oryza sativa, which are operably linked to heterologous polynucleotides in recombinant DNA constructs, to regulate gene expression in plants, ensuring optimal expression and stability of transgenes by functioning as transcription termination signals.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple genes are transferred to create transgenic plants with multiple traits, then the plant can express complex phenotypes, but repetitive sequences can cause undesirable negative effects on transgene expression and stability
Solution Approach 1:
The patent divides the regulatory region into distinct functional segments (promoters, terminators, enhancers, silencers) that can be independently selected and combined. This segmentation allows optimization of each element separately to avoid repetitive sequence issues while maintaining multiple trait expression capability.
Solution Approach 2:
The patent applies different regulatory elements with specific local functions to different transgenes. Each transgene can be equipped with tailored regulatory regions (unique promoters, terminators, enhancers) that are optimized for its specific expression requirements, preventing the negative effects of repetitive sequences while maintaining versatility.
2Manufacturing precision
If diverse regulatory elements are used to control each transgene optimally, then transgene expression can be modulated precisely, but the complexity of the recombinant DNA construct increases
Solution Approach 1:
The patent identifies and utilizes universal regulatory elements that can function across multiple transgenes and plant species. Standardized promoters, terminators, enhancers, and silencers can be reused in different combinations, providing precise expression control without proportionally increasing construct complexity.
Solution Approach 2:
The patent allows for selective inclusion of regulatory elements based on specific expression needs. Not all transgenes require all types of regulatory elements; the construct complexity is optimized by including only the necessary promoters, terminators, enhancers, or silencers for each specific application.
3Manufacturing precision
If terminator sequences are used to regulate gene expression, then transcription can be terminated precisely, but improper termination can lead to read-through and affect mRNA stability
Solution Approach 1:
The patent utilizes terminator sequences as intermediary elements between the coding region and downstream sequences. These terminators act as mediators that precisely control transcription termination and facilitate proper 3' mRNA processing, including polyadenylation, thereby ensuring both accurate termination and mRNA stability without read-through effects.
Data Source
AI summary
The present disclosure discloses polynucleotide sequences that can be used to regulate gene expression in plants. Terminator sequences from Sorghum bicolor and Oryza sativa that are functional in plants are disclosed. Nucleic acid molecules, recombinant expression constructs, plants and seed comprising these terminator sequences are further disclosed.