CRISPR-Cas SGR Gene Editing for Soybean Rust Resistance

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Solution Overview

Problem

Current genetic resistance mechanisms for soybean rust, such as Rpp genes, are not durable due to rapid evolution of virulence in pathogens, and fungicides become less effective over time, necessitating a novel strategy for improved resistance.

Innovation Solution

Introduction of non-natural mutations in the STAYGREEN (SGR) gene using a CRISPR-Cas editing system to enhance resistance to soybean rust in soybean plants, specifically through targeted editing of the SGR gene to introduce substitutions, insertions, or deletions, resulting in transgene-free edited plants with increased resistance.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If Rpp genes are used for soybean rust resistance, then race-specific resistance is provided, but resistance breakdown occurs rapidly due to pathogen evolution

Engineering Contradiction:

Improvedurability of resistanceVSAvoidpathogen virulence evolution

Core Design Contradiction:

ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent modifies the SGR gene sequence to create non-natural mutations that alter the gene's function or expression level, thereby changing the plant's physiological parameters to achieve durable resistance against soybean rust without relying on race-specific Rpp genes that are quickly overcome by pathogen evolution

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

Instead of using traditional resistance genes (Rpp) that directly对抗 the pathogen, the patent inverts the approach by modifying a gene (SGR) involved in chlorophyll metabolism to indirectly confer resistance, creating a novel resistance mechanism that does not follow the conventional path of direct pathogen-plant interaction

Inventive Principle:

Principle #13The other way round (Inversion)

2Object-affected harmful factors

If fungicides are used for soybean rust control, then disease suppression is achieved, but effectiveness decreases over time due to resistance development

Engineering Contradiction:

Improvesoybean rust suppressionVSAvoidfungicide effectiveness duration

Core Design Contradiction:

Object-affected harmful factorsVSDuration of action of stationary object

Solution Approach 1:

The patent converts the harmful effect of pathogen evolution and fungicide resistance into a benefit by developing a genetic resistance mechanism that is not subject to the same resistance breakdown, using SGR gene modification to create lasting protection that improves upon the temporary suppression provided by fungicides

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

3Ease of manufacture

If CRISPR-Cas editing is used to modify SGR gene, then transgene-free edited plants are produced, but editing precision and off-target effects must be managed

Engineering Contradiction:

Improvetransgene-free plant productionVSAvoidgene editing accuracy

Core Design Contradiction:

Ease of manufactureVSManufacturing precision

Solution Approach 1:

The patent uses guide RNA as an intermediary that directs the Cas9 enzyme to the specific target site in the SGR gene, ensuring precise editing while allowing the system to be delivered and expressed in the plant cells, ultimately producing transgene-free edited plants with high accuracy

Inventive Principle:

Principle #24Intermediary (Mediator)

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 method generates soybean plants with increased resistance to soybean rust, providing a durable solution by modifying the SGR gene using CRISPR-Cas technology, which is transgene-free and effective against multiple isolates of the pathogen.

Implementation Method 1

a CRISPR-Cas effector protein; and a guide nucleic acid (gRNA, gDNA, crRNA, crDNA, sgRNA, sgDNA) comprising a spacer sequence with complementarity to an endogenous target gene encoding a SGR protein

Methodology Applied

Scientific EffectCRISPR-Cas gene editing:

Data Source

PatentUS20240292803A1Methods for improving resistance to soybean rust

Publication Date: 2024.09.05 PAIRWISE PLANTS SERVICES INC

AI summary

This invention relates to compositions and methods for modifying STAYGREEN (SGR) genes in plants. The invention further relates to plants produced using the methods and compositions of the invention comprising modified endogenous SGR genes and having increased resistance to soybean rust.