Genetically Modified Transformant for Phosphite-Based Containment

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Solution Overview

Problem

Current methods for preparing genetically modified organisms (GMOs) with high containment effects are complex and require introducing defects into multiple genes, making them difficult to prepare and maintain.

Innovation Solution

A transformant is developed that is defective in phosphate and phosphate ester transporter functions but capable of utilizing reduced phosphorous compounds like phosphite for proliferation, achieved by introducing a hypophosphite transporter protein, allowing for simpler preparation and high containment.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If multiple gene defects are introduced to achieve high containment effect, then containment reliability is improved, but device complexity increases

Engineering Contradiction:
Improvecontainment effectVSAvoidpreparation complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent extracts the essential function of containment by focusing on a single critical dependency - the ability to utilize reduced phosphorous compounds. Instead of modifying multiple genes across the genome, the invention identifies and modifies only the phosphorous metabolism pathway, specifically introducing a hypophosphite transporter protein while deleting phosphate transporter functions. This extraction of the critical function simplifies the overall system while maintaining containment reliability.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent changes the metabolic parameter of phosphorous utilization by introducing a hypophosphite transporter protein that enables uptake of reduced phosphorous compounds. This parameter change transforms the organism's nutritional requirement from natural phosphate to synthetic reduced phosphorous compounds, creating a containment mechanism that does not require multiple gene defects. The single parameter change in phosphorous metabolism achieves the same containment effect as complex multi-gene modifications.

Inventive Principle:
Principle #35Parameter changes

2Reliability

If multiple gene defects are introduced to achieve high containment effect, then containment reliability is improved, but manufacturing precision deteriorates

Engineering Contradiction:
Improvecontainment effectVSAvoidpreparation precision
Core Design Contradiction:
ReliabilityVSManufacturing precision

Solution Approach 1:

The patent extracts the essential function of containment by focusing on a single critical dependency - the ability to utilize reduced phosphorous compounds. Instead of modifying multiple genes across the genome, the invention identifies and modifies only the phosphorous metabolism pathway, specifically introducing a hypophosphite transporter protein while deleting phosphate transporter functions. This extraction of the critical function simplifies the overall system while maintaining containment reliability.

Inventive Principle:
Principle #2Taking out (Extraction)

3Ease of operation

If natural phosphate is used for proliferation, then ease of operation is improved, but containment reliability deteriorates

Engineering Contradiction:
Improveculture medium availabilityVSAvoidcontainment effect
Core Design Contradiction:
Ease of operationVSReliability

Solution Approach 1:

The patent inverts the conventional approach by not using natural phosphate as the growth substrate but rather requiring synthetic reduced phosphorous compounds. This inversion creates a containment mechanism where the organism cannot survive in natural environments lacking these synthetic compounds. The hypophosphite transporter protein enables uptake of reduced phosphorous compounds, transforming the organism's nutritional requirement from natural to synthetic sources, thereby achieving both ease of operation in controlled settings and high containment reliability.

Inventive Principle:
Principle #13The other way round (Inversion)

4Ease of operation

If antibiotics are used for selection, then ease of operation is improved, but loss of substance increases

Engineering Contradiction:
Improveselection methodVSAvoidcost reduction
Core Design Contradiction:
Ease of operationVSLoss of substance

Solution Approach 1:

The patent replaces expensive, persistent antibiotics with a cheaper, biodegradable selection mechanism based on reduced phosphorous compound utilization. The hypophosphite transporter protein enables the organism to grow on inexpensive reduced phosphorous compounds that do not require antibiotic selection. This substitution eliminates the need for continuous antibiotic presence, reducing substance loss and cost while maintaining ease of operation through simple media composition changes.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

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 transformant can be easily prepared using a small number of genetic manipulations, provides a high containment effect by relying on non-natural phosphorous compounds, and reduces costs by using inexpensive culture media, avoiding the need for antibiotics.

Implementation Method 1

a gene encoding a hypophosphite transporter protein is introduced... the hypophosphite transporter protein having a function to transport a reduced phosphorous compound to cells

Methodology Applied
Scientific EffectActive transport:

Implementation Method 2

the transformant being incapable of utilizing phosphate for proliferation but capable of utilizing phosphite for proliferation

Methodology Applied
Scientific EffectOxidation: Oxidation

Data Source

PatentEP3508575B1Transformant, method for producing transformant, and method for detecting presence or absence of reduced phosphorus compound using transformant
Publication Date: 2020.12.16 HIROSHIMA UNIVERSITY
  • EP3508575B1 patent drawingFigure 1
  • EP3508575B1 patent drawingFigure 2
  • EP3508575B1 patent drawingFigure 3

AI summary

In order to provide a transformant by which a high containment effect is obtained and which can be prepared by a simple procedure, a method for producing the transformant, and a method for detecting the presence of a reduced phosphorous compound with use of the transformant, a transformant in accordance with an embodiment of the present invention is a transformant which is defective in functions of a gene encoding a phosphate transporter protein and a gene encoding a phosphate ester transporter protein and into which a gene encoding a hypophosphite transporter protein is introduced.