Gene-edited Soybean Reduces Isoflavone Content Without Transgenic DNA
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Solution Overview
Problem
Current methods for reducing isoflavone levels in soybeans, such as TILLING, transgenic overexpression, and suppression, fail to deliver products free of unintended mutations or transgenic DNA sequences, and do not meet consumer demand for non-transgenic soybean products with lower isoflavone content.
Innovation Solution
Targeted mutagenesis using genome editing techniques, specifically introducing mutations in the coding regions of IFS1, IFS2, Myb29, and their regulatory sequences, to reduce isoflavone expression in soybean plants without introducing transgenic DNA, utilizing rare-cutting endonucleases like TALENs to achieve specific reductions in isoflavone content.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If transgenic methods are used to reduce isoflavone levels, then isoflavone content is reduced, but transgenic DNA sequences are introduced which consumers reject
Solution Approach 1:
The patent extracts and removes the harmful transgenic DNA sequences from the system while retaining the desired effect of reduced isoflavone content. By using CRISPR/Cas9 gene editing, the invention targets specific genes (IFS1, IFS2, Myb29) for modification without introducing foreign transgenic DNA, thus taking out the harmful element while preserving the beneficial outcome.
Solution Approach 2:
The patent uses an intermediary approach by employing guide RNA and Cas9 enzyme as mediators to achieve precise gene editing. These intermediaries enable targeted modification of isoflavone biosynthesis genes without requiring transgenic DNA integration, thus reducing isoflavone content while avoiding the introduction of harmful transgenic sequences.
2Quantity of substance
If traditional breeding methods are used, then isoflavone levels may be reduced, but the process is time-consuming and less precise
Solution Approach 1:
The patent replaces the mechanical and time-consuming traditional breeding system with a molecular-level gene editing system. By using CRISPR/Cas9 technology, the invention directly modifies the DNA sequences of target genes (IFS1, IFS2, Myb29) to reduce isoflavone content, substituting the slow process of selective breeding with precise molecular manipulation that achieves the desired trait in a single generation.
Solution Approach 2:
The patent performs preliminary action by designing and introducing specific guide RNAs and Cas9 enzymes that target predetermined gene sequences before the breeding process begins. This pre-planned molecular intervention directly modifies the genes responsible for isoflavone synthesis, eliminating the need for multiple generations of selective breeding and significantly reducing the time required to achieve the desired isoflavone reduction.
3Quantity of substance
If TILLING or transgenic suppression methods are used, then isoflavone levels are reduced, but unintended mutations occur
Solution Approach 1:
The patent applies local quality by using highly specific guide RNAs that target only the precise locations of isoflavone biosynthesis genes (IFS1, IFS2, Myb29). The CRISPR/Cas9 system introduces mutations only at these specific target sites through programmed DNA cleavage and repair, ensuring that gene modification is localized exactly where needed while leaving the rest of the genome unchanged, thus achieving high mutation specificity.
Solution Approach 2:
The patent replaces the random or broad-spectrum mutation methods of TILLING with a precision molecular editing system. By using guide RNA-directed Cas9 cleavage, the invention substitutes random mutagenesis with targeted, site-specific gene modification, achieving precise reduction of isoflavone content while minimizing unintended mutations elsewhere in the genome.
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 effectively reduces isoflavone content in soybean plants by up to 99% without introducing transgenic sequences, meeting consumer demands for non-transgenic soybean products with lower isoflavone levels, and can be applied across various soybean varieties.
Implementation Method 1
one or more mutations is introduced using a targeted genome modification using a rare-cutting endonuclease
Implementation Method 2
the rare-cutting endonuclease is a transcription activator-like effector nuclease (TALEN)
Data Source
AI summary
Gene-edited soybean plants, plant parts, and plant cells that have a reduced isoflavone content compared to the corresponding unaltered soybean plant, plant part, or plant cell and methods of their production are disclosed. For example, introducing one or more mutations in the coding region of Myb29 gene, the MYBCORE binding motif of the IFS2 promoter, the MYBCORE binding motif of the IFS1 promoter, the coding region of the IFS2 gene, the coding region of the IFS1 gene, or a combination thereof is used to produce the gene-edited plant, plant part or plant cell via targeted genome modification.