Cytokinin Receptor HK Gene Editing for Fine-Tuned Yield Traits
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Solution Overview
Problem
Existing breeding and transgenic approaches have failed to achieve a significant step change in plant yield due to the complexity of yield traits, and modifying hormone pathways through transgene technology has not provided the necessary fine-tuning for yield improvement.
Innovation Solution
Introduce mutations in the Cytokinin Receptor Histidine Kinase (HK) genes in plants using CRISPR-Cas editing systems to generate specific edits in the HK gene sequences, thereby enhancing yield traits such as seed size and oil content without introducing transgenes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If transgenic approaches involving stable transformation are used to increase yield, then hormone related pathways can be modified, but the simple over- or under-expression through transgene technology is not consistent with the fine-tuning effect required for improving plant yield
Solution Approach 1:
The patent extracts and removes the transgene expression system, instead using CRISPR-Cas9 to directly edit the endogenous HK gene sequences. This eliminates the coarse over-expression mechanism of transgenes and enables precise, site-specific modifications to the native gene, achieving the required fine-tuning precision for yield improvement while maintaining the ability to modify hormone pathways.
Solution Approach 2:
The patent replaces the mechanical transgene expression system with a molecular editing system (CRISPR-Cas9). Instead of relying on transgene transcription and translation machinery to achieve hormone pathway modification, the system uses programmable nucleases to directly alter the genetic code of endogenous genes, enabling precise control over gene function and expression levels.
2Productivity
If intensive breeding is used to increase plant yield, then incremental increases in yield can be achieved, but genetic gain from breeding has started to plateau and assembling multiple small-effect genes has substantially increased research and development costs
Solution Approach 1:
The patent changes the fundamental parameter of genetic modification from assembling multiple small-effect genes through conventional breeding to targeting and modifying key regulatory genes (HK genes) using CRISPR-Cas9. This shifts the approach from quantitative accumulation of many minor effects to qualitative changes in critical control points, thereby achieving yield improvement without the complexity of managing multiple gene assemblies.
Solution Approach 2:
The patent extracts the complex breeding process of assembling multiple small-effect genes and replaces it with a targeted gene editing approach. By focusing on specific histidine kinase genes that regulate hormone pathways, the method eliminates the need to assemble numerous small-effect genes, reducing research and development costs while maintaining productivity gains.
3Manufacturing precision
If CRISPR-Cas editing systems are used to introduce mutations in HK genes, then specific edits can be generated to enhance yield traits, but the process requires precise targeting and selection of progeny plants
Solution Approach 1:
The patent performs preliminary action by designing and introducing guide RNAs and Cas9 proteins into the plant system before the actual gene editing occurs. The CRISPR-Cas9 system is pre-assembled and delivered into plant cells, where it autonomously identifies and edits the target HK genes. This preliminary setup enables precise editing without requiring time-consuming sequential steps during the actual modification process.
Solution Approach 2:
The CRISPR-Cas9 system is designed to be self-directed, where the guide RNA sequences autonomously direct the Cas9 nuclease to the correct target loci in the genome. The system self-identifies the target HK genes, performs the cutting, and enables repair through cellular mechanisms, reducing the need for extensive manual selection and screening of progeny plants while maintaining high precision.
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 produces plants with improved yield traits, including increased seed size and oil content, by precisely modifying the HK genes, overcoming the limitations of previous methods.
Implementation Method 1
A second aspect of the invention provides a plant cell, comprising an editing system comprising: (a) a CRISPR-Cas effector protein; and (b) a guide nucleic acid comprising a spacer sequence with complementarity to an endogenous target gene encoding a histidine kinase (HK) polypeptide.
Data Source
AI summary
This invention relates to compositions and methods for modifying Cytokinin Receptor Histidine Kinase (HK) genes in plants, optionally to improve yield traits, the HK genes encoding a histidine kinase (HK) polypeptide. The invention further relates to plants having increased improved yield traits produced using the methods and compositions of the invention.