RNA Aptamer Ligand-Controlled Gene Expression in Transgenic Plants
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for gene suppression in plants, such as anti-sense and RNA interference, have low efficiency and often result in unintentional systemic spreading of gene suppression, and there is a need for targeted and specific regulation of gene expression in plants and pests.
Innovation Solution
Development of transgenic plants with recombinant DNA constructs that include RNA aptamers capable of binding to specific ligands, which regulate gene expression through allosterically affected regulatory RNA domains, allowing for ligand-controlled expression or suppression of target sequences, and the use of introns to deliver gene suppression elements without protein-coding exons.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If anti-sense and RNA interference methods are used for gene suppression in plants, then gene expression can be suppressed, but the efficiency is low and unintentional systemic spreading occurs
Solution Approach 1:
The patent applies local quality by using tissue-specific promoters to drive RNA aptamer expression only in specific plant tissues or cells where pest or pathogen presence is detected. This localized expression approach ensures that gene suppression occurs only in the affected areas rather than systemically throughout the entire plant, thereby preventing unintentional systemic spreading while maintaining effective local control of gene expression for pest and pathogen management
Solution Approach 2:
The patent introduces RNA aptamers as intermediary molecules that specifically bind to ligands from pests or pathogens. These RNA aptamers act as mediators between the detection of pest/pathogen presence and the activation of gene suppression mechanisms. The aptamers translate the presence of specific ligands into conformational changes that regulate downstream gene expression, providing a targeted and controlled response rather than indiscriminate systemic suppression
2Adaptability or versatility
If transgenic plants with recombinant DNA constructs are developed, then precise control of gene expression is achieved, but device complexity increases
Solution Approach 1:
The patent employs dynamic control mechanisms where RNA aptamers undergo conformational changes in response to ligand binding events. These dynamic structural transitions in the RNA aptamers allow the system to switch between different states (bound/unbound, active/inactive) based on the presence or absence of specific ligands from pests or pathogens, enabling precise and adaptable gene expression control without requiring complex multi-component systems
Solution Approach 2:
The patent achieves versatility through the use of modular recombinant DNA constructs that can be adapted to target multiple different ligands by simply changing the RNA aptamer sequence while maintaining the same basic construct architecture. The universal promoter-aptamer-regulated gene framework can recognize and respond to diverse pest and pathogen ligands, providing multi-functional capability without proportionally increasing overall system complexity
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enables precise and efficient control of gene expression in plants and pests, reducing damage from invertebrate pests and pathogens, and avoids unintentional systemic spreading of gene suppression, improving the specificity and efficacy of gene regulation.
Implementation Method 1
Nucleic acid aptamers include DNA or RNA sequences that can recognize and specifically bind, often with high affinity, a particular molecule or ligand
Implementation Method 2
regulate expression of a target sequence, wherein the regulation of the target sequence is dependent on the conformation of the regulatory RNA, and the conformation of the regulatory RNA is allosterically affected by the binding state of the RNA aptamer
Data Source
AI summary
The present invention provides molecular constructs and methods for use thereof, including constructs including heterologous miRNA recognition sites, constructs for gene suppression including a gene suppression element embedded within an intron flanked on one or on both sides by non-protein-coding sequence, constructs containing engineered miRNA or miRNA precursors, and constructs for suppression of production of mature microRNA in a cell. Also provided are transgenic plant cells, plants, and seeds containing such constructs, and methods for their use. The invention further provides transgenic plant cells, plants, and seeds containing recombinant DNA for the ligand-controlled expression of a target sequence, which may be endogenous or exogenous. Also disclosed are novel miRNAs and miRNA precursors from crop plants including maize and soy.