Plant Resistance Suppression for High Target Protein Expression
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Solution Overview
Problem
Existing methods for producing target substances in plants face challenges due to activation of self-defense mechanisms that suppress the expression of exogenous genes, leading to reduced production of desired proteins.
Innovation Solution
A method is developed to suppress or disrupt genes associated with resistance mechanisms in plants, such as those in the salicylic acid (SA) biosynthetic pathway and signal transduction, or overexpress genes related to SA degradation, using nucleic acid constructs and transient expression systems like agroinfiltration and plant viral vectors, to create plants with suppressed resistance and high target protein expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If exogenous genes are transferred into plants to produce desired proteins, then target substance production is enabled, but plant self-defense mechanisms are activated which suppress exogenous gene expression, reducing production efficiency
Solution Approach 1:
The patent applies preliminary anti-action by suppressing plant resistance mechanisms before introducing the exogenous gene. Specifically, resistance-suppressing genes (such as those encoding protease inhibitors or RNAi components) are introduced first to disable the plant's defense system, thereby preventing subsequent suppression of the target gene expression and ensuring high productivity.
Solution Approach 2:
Instead of trying to enhance plant resistance while producing target substances, the patent inverts the approach by deliberately suppressing resistance mechanisms. This inversion allows the plant to become more susceptible to the exogenous gene expression, thereby resolving the contradiction between maintaining resistance and achieving high target protein production.
2Reliability
If plant resistance mechanisms are reinforced to protect against pathogens, then pathogen resistance is improved, but expression of exogenous genes is suppressed, lowering target substance production
Solution Approach 1:
The patent inverts the conventional approach by suppressing resistance mechanisms rather than reinforcing them. This allows exogenous genes to be expressed at high levels for target substance production, while the plant's susceptibility to pathogens is temporarily increased as a trade-off.
Solution Approach 2:
The patent changes the parameter of resistance level from high to low by introducing resistance-suppressing genes. This parameter change enables high-level expression of exogenous genes, thereby achieving high productivity for target substance production, while accepting reduced pathogen resistance as a consequence.
3Ease of manufacture
If transient expression systems utilize pathogen infection mechanisms for gene transfer, then gene delivery efficiency is improved, but activation of self-defense mechanisms suppresses exogenous gene expression, reducing target protein production
Solution Approach 1:
The patent applies preliminary anti-action by introducing resistance-suppressing genes before using transient expression systems for gene transfer. This preliminary action disables the plant's defense system that would otherwise be activated by the infection mechanism, thereby ensuring that the exogenous gene is expressed at high levels and target protein production is maximized.
Solution Approach 2:
The patent performs preliminary action by pre-suppressing resistance mechanisms through introduction of resistance-suppressing genes. This preliminary suppression creates a favorable environment for subsequent transient expression of exogenous genes, ensuring high target protein production while utilizing efficient gene transfer methods.
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 allows for high expression of target proteins in plants by inhibiting their resistance mechanisms, enabling effective production of transformed or genome-edited plants with suppressed resistance to pathogens.
Implementation Method 1
NPL 1 describes creating recombinant plant species with suppression of genes related to gene silencing, which is a major function among plant resistance mechanisms
Implementation Method 2
using nucleic acid constructs and transient expression systems like agroinfiltration and plant viral vectors
Implementation Method 3
using nucleic acid constructs and transient expression systems like agroinfiltration and plant viral vectors
Data Source
AI summary
Provided is a method for achieving high expression of a target substance in a plant. Also provided are a method of creating a plant with suppressed resistance to a pathogen, wherein the method comprises suppression or disruption of expression of a gene associated with a resistance mechanism in the plant, and a method of creating a plant with suppressed resistance to a pathogen wherein the method comprises overexpression of a gene related to degradation of salicylic acid (SA) in plants; as well as a transgenic plant or genome-edited plant obtained by the method; and a method for producing a target protein using the plant.