Mutant GBSS1 Polypeptides for Precise Amylose Content Control
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Solution Overview
Problem
Current methods are inadequate for precisely modifying the amylose content (AC) in rice using CRISPR/Cas editing technology to improve cooking quality, as they face challenges in efficiently altering the Waxy gene to achieve desired AC levels.
Innovation Solution
Development of mutant GBSS1 polypeptides with specific amino acid mutations, such as at positions 168, 237, 411, and 428, to alter the amylose content in plants, utilizing CRISPR/Cas editing to introduce targeted mutations into the GBSS1 gene, thereby modifying the AC in rice.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If CRISPR/Cas editing technology is used to modify the Waxy gene to change amylose content, then the cooking quality of rice can be improved, but the efficiency and precision of altering the GBSS1 gene to achieve desired AC levels is insufficient
Solution Approach 1:
The patent applies parameter changes by identifying specific amino acid positions (168, 237, 411, 428) in the GBSS1 protein that, when mutated, directly alter the enzyme's activity and thereby control amylose content. This targeted approach to changing protein parameters enables precise control over AC levels while improving editing efficiency through focused mutagenesis at these critical positions.
2Adaptability or versatility
If natural allelic variation in the Waxy locus is used to control amylose synthesis, then amylose content can be naturally varied, but the precision and range of AC modification is limited
Solution Approach 1:
The patent applies local quality by focusing mutations at specific local positions (amino acid residues 168, 237, 411, 428) within the GBSS1 protein structure. These localized changes at critical functional positions enable precise control over amylose synthesis, allowing researchers to achieve desired AC levels with high precision while maintaining the overall protein structure and function.
Data Source
AI summary
A mutant granule-bound starch synthase 1 (GBSS1) polypeptide and a nucleic acid, and use thereof are provided. Compared to an amino acid sequence of a parent GBSS1, the mutant GBSS1 polypeptide has a mutation at an amino acid corresponding to one or more of amino acid 237, amino acid 168, and amino acid 411 of an amino acid sequence shown in SEQ ID NO: 1. An amylose content (AC) in a plant changes after the plant undergoes GBSS1 mutation, which has very promising application prospects in the improvement of edible quality of rice.