Plant Terminator Sequences for Bidirectional Transcription Control
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Solution Overview
Problem
Existing plant genetic engineering techniques face challenges in optimally modulating the expression of multiple transgenes due to issues like read-through transcription, leaky terminators, and convergent gene transcription, leading to undesirable effects on transgene stability and expression.
Innovation Solution
The development of novel plant terminator sequences, including specific nucleotide sequences and their functional fragments, which act as efficient transcriptional and bidirectional terminators to regulate gene expression, ensuring optimal expression of heterologous polynucleotides in plants.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional terminator sequences are used in transgenic plants, then transcription termination is achieved, but read-through transcription and leaky termination occur leading to unwanted gene expression
Solution Approach 1:
The patent modifies terminator sequence parameters by introducing mutated versions with altered nucleotide compositions while maintaining functional elements. The mutated terminators show improved termination efficiency and reduced read-through transcription compared to conventional sequences, demonstrating parameter optimization to resolve the contradiction between reliable termination and harmful read-through effects
Solution Approach 2:
The invention creates composite regulatory elements by combining promoter regions, terminator sequences, and mutated terminal repeats into integrated recombinant DNA constructs. These composite structures work synergistically to achieve robust transcription termination while preventing leaky expression and unwanted gene overlaps
2Adaptability or versatility
If multiple transgenes are introduced into plants for gene stacking, then improved characteristics are achieved, but repetitive sequences cause negative effects on transgene expression and stability
Solution Approach 1:
The patent applies local quality by using distinct regulatory elements for each transgene within the stacking construct. Each gene is paired with its own unique promoter and terminator combination, preventing repetitive sequence issues while maintaining stable expression of multiple traits. This localized differentiation resolves the contradiction between multi-trait capability and genomic stability
3Quantity of substance
If convergent transcription of transgenes occurs, then gene density is increased, but overlapping transcription decreases expression and generates antisense RNA
Solution Approach 1:
The patent introduces bidirectional terminators as intermediary elements between convergently oriented transgenes. These specialized terminator sequences actively prevent transcriptional interference and antisense RNA generation, mediating the interaction between adjacent genes to maintain stable expression while allowing high gene density in the construct
Data Source
AI summary
This invention relates to gene expression regulatory sequences, specifically transcription terminator sequences. Plant transcription terminator sequences are described herein. Methods for identifying novel plant transcription terminator sequences that can terminate transcription in one orientation or in a bidirectional manner and methods of using these terminator sequences to generate transgenic plants are described herein.