Transient Morphoregulator Activation for Plant Transformation
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Solution Overview
Problem
Current methods for genetic transformation in plants, particularly recalcitrant species like corn, rely on transgenic approaches that involve genomic insertion of morphoregulator-encoding transgenes, which are restricted by regulations and require complex segregation processes, lengthening development and production timelines and increasing costs.
Innovation Solution
Transiently increasing the expression of endogenous ODP2 and/or WUS2 polypeptides in plant cells using exogenous gene transcription agents that stimulate transcription, allowing for the formation of regenerable plant structures without the need for genomic insertion, thereby facilitating genetic editing and transformation across various plant species.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If transgenes encoding morphoregulators are genomically inserted to stimulate somatic embryogenesis, then plant regenerative potential is improved, but regulatory restrictions and complex segregation processes increase development time and costs
Solution Approach 1:
The patent uses an exogenous transcription agent as an intermediary to temporarily activate endogenous morphoregulator genes (ODP2 and/or WUS2) without genomic insertion. This mediator stimulates somatic embryogenesis and regenerative potential while avoiding the regulatory and temporal drawbacks of transgenic approaches. The transcription agent acts as a temporary catalyst that can be removed, leaving no permanent transgenic modification.
Solution Approach 2:
The patent extracts the essential function of morphoregulator transgenes (stimulating somatic embryogenesis) and achieves it through transient activation of endogenous genes rather than permanent genomic insertion. By taking out the transgenic component and using only the functional effect temporarily, the method avoids regulatory restrictions and segregation requirements while maintaining the desired regenerative outcome.
2Productivity
If transgenes are genomically inserted to enable selectable marker enrichment, then genetic editing efficiency is improved, but segregation processes and multiple crosses are required
Solution Approach 1:
The patent extracts the beneficial effect of selectable markers (enrichment for edited tissue) without requiring permanent transgenic insertion. By using transient transcriptional activation of endogenous genes, the method achieves tissue enrichment and selection capability while avoiding the need for segregating transgenic lines through multiple crosses.
Solution Approach 2:
The patent employs a disposable, transient transcription agent that provides selectable marker functionality only during the editing process. This short-lived agent enables efficient selection of edited cells without creating permanent transgenic lines that would require complex segregation and multiple crossing generations to eliminate.
3Adaptability or versatility
If morphoregulator transgenes are used to make recalcitrant tissues accessible to transformation, then transformation accessibility is improved, but genomic insertion restrictions apply
Solution Approach 1:
The patent uses an exogenous transcription agent as a temporary intermediary to activate endogenous morphoregulators in recalcitrant tissues, making them accessible to transformation. This approach provides the versatility of transgenic activation while maintaining regulatory compliance by avoiding permanent genomic insertion, thus easing manufacturing constraints.
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 enhances plant regenerative potential, improves genetic editing efficiency, and reduces regulatory and production hurdles by avoiding the need for genomic insertion, enabling faster and more cost-effective production of genetically edited plants.
Implementation Method 1
the expression of the endogenous ODP2 polypeptide and/or the endogenous WUS2 polypeptide is transiently increased in the plant cell with at least one exogenous gene transcription agent that stimulates transcription of the endogenous ODP2 gene and/or with at least one exogenous gene transcription agent that stimulates transcription of the endogenous WUS2 gene
Data Source
AI summary
Plant cells and related systems, methods, and compositions for improving the capacity of the plant cells to regenerate embryogenic plant tissues, plant organs, and whole plants are provided. Such plant cells and related systems, methods, and compositions provide for increased expression of the endogenous morphoregulators BABYBOOM (ODP2) and/or WUSCHEL2 (WUS2).