Wheat SBEII Gene Knockout for Resistant Starch
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Solution Overview
Problem
Current wheat varieties have a starch composition that is predominantly amylopectin, making them quickly digestible and potentially contributing to diet-related chronic diseases. There is a need for wheat varieties with increased amylose content, resistant starch, and dietary fiber to improve nutritional quality.
Innovation Solution
The use of sequence-specific nucleases, such as TALE nucleases, to introduce targeted mutations or knockouts in the SBEII genes, specifically in the SBEIIa and SBEIIb alleles, to reduce the expression and activity of starch-branching enzymes, thereby increasing the amylose content in wheat grains.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If sequence-specific nucleases are used to knockout SBEII genes, then amylose content and dietary fiber are increased, but the complexity of the genetic modification process increases
Solution Approach 1:
The patent replaces traditional mechanical breeding methods with sequence-specific nucleases (molecular tools) to achieve precise gene knockout. This substitution enables targeted modification of SBEII genes to increase amylose content without the randomness and complexity of conventional breeding approaches.
Solution Approach 2:
The patent extracts and removes the harmful or limiting factor (SBEII genes responsible for amylopectin synthesis) through targeted knockout. By eliminating these specific genes, the natural balance shifts toward increased amylose production, achieving the desired compositional change.
2Quantity of substance
If SBEII genes are knocked out to increase amylose, then nutritional quality improves, but the difficulty of detecting and measuring gene modifications increases
Solution Approach 1:
The patent employs feedback mechanisms through molecular markers and detection systems that allow researchers to verify successful gene knockout and track the desired phenotypic changes. This feedback enables confirmation that SBEII genes have been effectively disabled and that amylose content has increased as intended.
3Quantity of substance
If targeted mutations are introduced in SBEII alleles, then resistant starch increases, but the manufacturing precision requirements increase
Solution Approach 1:
The patent applies local quality by introducing mutations at specific locations within the SBEII gene sequences. Rather than random mutagenesis, the sequence-specific nucleases target precise genomic locations to disable gene function, ensuring the mutation occurs where it will effectively reduce enzyme activity and increase resistant starch.
Solution Approach 2:
The patent replaces imprecise traditional mutagenesis methods with sequence-specific molecular tools that can accurately target and modify specific gene sequences. This substitution achieves the required manufacturing precision for targeted mutations, ensuring modifications occur only at the intended SBEII gene loci.
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 results in wheat varieties with significantly higher levels of amylose, resistant starch, and dietary fiber, potentially leading to improved nutritional qualities and health benefits, such as slower glucose absorption and reduced risk of chronic diseases.
Implementation Method 1
The TALE nuclease can bind to a sequence as set forth in SEQ ID NO: 1
Implementation Method 2
sequence-specific nucleases to make targeted mutations or knockouts of the SBEII genes
Data Source
AI summary
Materials and methods are provided for making plants (e.g., Triticum varieties) with increased levels of dietary fiber, such as by making TALE nuclease-induced mutations in alleles encoding starch branching enzyme IIa (SBEIIa) and starch branching enzyme IIb (SBEIIb).