CRISPR-Edited Sorghum IAP Locus for Cross-Compatibility
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Solution Overview
Problem
Sorghum bicolor's natural molecular mechanisms controlling cross-compatibility with its relatives hinder biotechnological and breeding developments due to cross-incompatibility issues, limiting the introduction of desirable stress-tolerance genes and gene flow between Sorghum species.
Innovation Solution
Genetically modified Sorghum plants are developed using the CRISPR/Cas9 gene editing system to modify genes in the inhibition of alien pollen (IAP) locus, enhancing or reducing cross-compatibility by knocking out or altering the expression of specific genes such as Sobic.002G023300, Sobic.002G023500, Sobic.002G023600, and Sobic.002G023700, allowing for improved compatibility with other Sorghum species and introducing stress-tolerance traits.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If natural cross-incompatibility mechanisms are maintained, then transgene escape to weed gene pools is prevented, but biotechnological development and gene flow between Sorghum species are limited
Solution Approach 1:
The patent modifies the IAP locus by introducing point mutations (e.g., G to A transitions) that change the genetic parameters of the cross-incompatibility mechanism. These parameter changes allow controlled adjustment of cross-compatibility levels, enabling both transgene containment and species cross-breeding depending on the specific mutation introduced.
Solution Approach 2:
Instead of maintaining the natural state of cross-incompatibility to prevent transgene escape, the patent inverts the approach by creating modified IAP alleles that are cross-compatible. This inversion allows transgenes to be intentionally transferred between species through controlled hybridization, while still maintaining containment through other mechanisms.
2Object-affected harmful factors
If cross-incompatibility is maintained to prevent unwanted gene flow, then breeding developments are restricted, but the risk of transgene escape to noxious weeds like Johnsongrass is reduced
Solution Approach 1:
The patent segments the IAP locus into multiple alleles with different compatibility characteristics (IAP1, IAP2, IAP3, etc.). Each allele provides a different level of cross-compatibility, allowing breeders to select specific combinations that balance transgene containment with breeding objectives. This segmentation enables fine-tuned control over gene flow.
Solution Approach 2:
The modified IAP alleles act as intermediaries that mediate between the conflicting requirements of transgene containment and breeding development. By introducing specific mutations in the IAP locus, the patent creates intermediate states of cross-compatibility that allow controlled gene flow for breeding purposes while maintaining sufficient barriers to prevent uncontrolled transgene escape to weeds.
3Reliability
If distant relatives' pollen tube growth is blocked by the IAP locus, then cross-incompatibility with species like Sorghum plumosum is maintained, but introduction of stress-tolerance genes is limited
Solution Approach 1:
The patent introduces specific parameter changes in the IAP locus genes (Sobic.002G023300, Sobic.002G023500, Sobic.002G023600) that modify pollen-pistil recognition parameters. These changes allow pollen from distant relatives like Sorghum plumosum to successfully grow through the pistil, enabling the introduction of stress-tolerance genes while maintaining controlled cross-compatibility.
Data Source
AI summary
The present disclosure provides genetically edited or modified plants of the genus Sorghum and methods of producing genetically edited or modified plants of the genus Sorghum using, e.g., a CRISPR/Cas9 gene editing tool. In some embodiments, the genetic modifications are directed to genes of the Inhibition of Alien Pollen (IAP) locus to modulate cross-compatibility of the plants with other species of Sorghum.