CRISPR-Edited Soybeans with Reduced KTI Activity

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Solution Overview

Problem

Soybean meal digestibility is impaired by trypsin inhibitors, requiring energy-intensive and costly heat treatment, which degrades essential amino acids, and breeding low trypsin inhibitor soybean cultivars is hindered by multiple TI genes and lack of molecular markers.

Innovation Solution

Engineered soybean plants with modified Kunitz trypsin inhibitor (KTI) genes, such as KTI1 and KTI3, using CRISPR-Cas mediated gene editing to reduce or eliminate trypsin inhibitor expression and activity, resulting in soybeans with improved nutritional profile.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Object-affected harmful factors

If heat treatment is applied to inactivate trypsin inhibitors, then trypsin inhibitor activity is reduced, but energy consumption increases and essential amino acids are degraded

Engineering Contradiction:

Improvetrypsin inhibitor activityVSAvoidenergy consumption

Core Design Contradiction:

Object-affected harmful factorsVSUse of energy by moving object

Solution Approach 1:

The patent applies preliminary action by genetically modifying the soybean plants during breeding to reduce or eliminate trypsin inhibitor expression before the seeds are even produced. This is achieved through CRISPR-Cas9 gene editing or traditional breeding of accessions with natural TI mutations, so that the resulting soybean varieties inherently have low TI activity without requiring subsequent heat treatment, thereby avoiding energy consumption and amino acid degradation

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent applies the extraction principle by specifically targeting and removing or disabling the trypsin inhibitor genes (Kunitz TI genes) from the soybean genome. Through gene editing techniques or selective breeding, the harmful TI components are extracted or eliminated from the soybean system, leaving the beneficial nutritional components intact without the need for energy-intensive processing

Inventive Principle:

Principle #2Taking out (Extraction)

2Object-affected harmful factors

If heat treatment is applied to inactivate trypsin inhibitors, then trypsin inhibitor activity is reduced, but essential amino acids are degraded

Engineering Contradiction:

Improvetrypsin inhibitor activityVSAvoidessential amino acids

Core Design Contradiction:

Object-affected harmful factorsVSLoss of substance

Solution Approach 1:

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent converts the natural genetic variation found in certain soybean accessions (which have reduced TI activity) into a beneficial trait through selective breeding. By identifying and propagating these natural mutants, the harmful TI effect is transformed into a benefit (low TI activity) while preserving the full amino acid profile, eliminating the need for heat treatment that would otherwise cause amino acid loss

Inventive Principle:

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

3Adaptability or versatility

If multiple members of TI genes are present, then natural genetic variation is available, but breeding success of low TI cultivars is hindered

Engineering Contradiction:

Improvenatural genetic variationVSAvoidbreeding complexity

Core Design Contradiction:

Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent introduces molecular markers as intermediary tools that bridge the gap between natural genetic variation and breeding selection. These markers serve as detectable indicators linked to low-TI alleles, allowing breeders to screen and select desirable plants without having to analyze complex multi-gene TI profiles, thus simplifying the breeding process while utilizing natural genetic diversity

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent replaces traditional mechanical/phenotypic selection methods with molecular marker-based selection. Instead of relying on complex phenotypic analysis or trial-and-error breeding approaches, the use of molecular markers provides a precise, efficient, and scalable method to identify and propagate low-TI variants, transforming the breeding system from a complex manual process to a streamlined molecular-guided process

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

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 engineered soybeans exhibit reduced trypsin inhibitor levels, enhancing protein digestibility and nutritional value, and provide biomarkers for plant selection, thus overcoming the limitations of current breeding methods.

Implementation Method 1

using CRISPR-Cas mediated gene editing to reduce or eliminate trypsin inhibitor expression and activity

Methodology Applied

Scientific EffectCRISPR-Cas mediated gene editing:

Data Source

PatentUS20230323375A1Soybeans having low trypsin inhibitor expression or activity

Publication Date: 2023.10.12 VIRGINIA TECH INTELLECTUAL PROPERTIES INC

AI summary

Described in several example embodiments herein are soybean plants and soybeans having one or more KTI genes having reduced or eliminated expression and/or activity thereby resulting soybean plants and soybeans having reduced or eliminated trypsin inhibition. In some embodiments, the soybean plants and soybeans are engineered to have one or more modified KTI genes. Also described herein are methods of making, growing, and using the soybean plants and soybeans described herein.