Self-Replicating Recombinant RNA for Precise Plant Phenotype Modification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
There is a need for modifying polynucleotides to improve phenotypes and genotypes of organisms, particularly in agricultural applications for crop plants.
Innovation Solution
A recombinant DNA molecule is developed, comprising a heterologous promoter operably linked to a DNA sequence encoding an RNA molecule with specific replicase recognition sequences for endornaviral RNA-dependent RNA polymerase (RDRP), a cargo RNA sequence, and optional additional elements such as viral movement proteins and encapsidation recognition sequences, to facilitate genetic modification and expression in plant cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional transformation methods are used to modify plants, then genetic modification can be achieved, but the efficiency and precision of phenotype improvement is limited
Solution Approach 1:
The invention divides the transformation system into separate functional modules: a DNA molecule encoding RNA, an RNA-dependent RNA polymerase (RdRP) from endornavirus, and specific RNA structures (5' and 3' replicase recognition sequences). This segmentation allows each component to be optimized independently and assembled for high-efficiency transformation with precise phenotypic outcomes.
Solution Approach 2:
The invention introduces an RNA intermediate molecule as a mediator between the DNA transformant and the final phenotypic change. The DNA encodes RNA that is processed by RdRP to produce functional RNA molecules that directly affect plant phenotypes, providing a precise and efficient transformation pathway.
2Productivity
If conventional RNA expression systems are used, then gene expression can be achieved, but replication efficiency and expression levels are insufficient
Solution Approach 1:
The invention employs self-replicating RNA molecules that contain 5' and 3' replicase recognition sequences. These sequences enable the RNA to direct its own replication through the endornavirus RdRP, achieving high replication efficiency without requiring complex external replication machinery. The RNA molecule serves its own replication needs through intrinsic structural elements.
Solution Approach 2:
The endornavirus RdRP serves multiple functions: it recognizes specific RNA structures, catalyzes RNA replication, and enables high-level gene expression. This multi-functional enzyme simplifies the overall expression system while achieving superior replication efficiency compared to conventional systems.
3Reliability
If standard genetic modification approaches are applied, then plant genotypes can be modified, but resistance to pests and pathogens is not sufficiently enhanced
Solution Approach 1:
The invention changes the fundamental parameters of genetic modification by using RNA-based transformation instead of traditional DNA integration methods. The RNA molecules produced by the endornavirus system directly modulate gene expression to enhance pest and pathogen resistance, providing reliable protection while maintaining relatively simple modification procedures.
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
The recombinant RNA molecule enables efficient replication and expression of cargo RNA, leading to phenotypic changes in plants, including increased resistance to pests and pathogens, stress tolerance, and targeted gene expression modulation.
Implementation Method 1
a 5' replicase recognition sequence that is capable of being recognized by an endornaviral RNA-dependent RNA polymerase (RDRP)... a 3' replicase recognition sequence that is capable of being recognized by the endornaviral RDRP
Data Source
AI summary
Synthetic endornaviral satellite RNA molecules and satellite particles containing the same are disclosed. The synthetic endornaviral satellite RNA molecules can include coding and/or non-coding cargo sequences, and are heritable through generations of plants. Also disclosed are methods of using the endornaviral satellite RNA molecules and satellite particles containing the same to change plant phenotypes, improve plant stress resistance, and improve plant pest and pathogen resistance.