Directed Recombination in Plant Breeding Using Custom Endonucleases
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Solution Overview
Problem
Traditional plant breeding methods rely heavily on visual observations and performance data, limiting the efficiency and precision of selecting optimal genetic traits, especially in hybrid crops, as they are constrained by the number and location of recombination events in breeding crosses.
Innovation Solution
The method involves identifying specific loci in a plant DNA sequence, introducing custom endonucleases to modify these loci, and using genetic tools to inhibit recombination or meiosis, allowing for precise genetic modifications such as targeted insertion, deletion, or disruption of genes, thereby enhancing breeding efficiency and trait selection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional plant breeding methods relying on visual observations and performance data are used, then the breeding process is simple and easy to operation, but the selection precision and efficiency of genetic traits is limited
Solution Approach 1:
The patent replaces traditional visual observation and phenotypic selection methods with molecular breeding techniques including DNA markers, genotyping, and molecular assays. This substitution enables precise identification of genetic traits at the molecular level, dramatically improving selection precision while the systematic approach manages the increased complexity through standardized protocols
Solution Approach 2:
The patent introduces molecular markers and DNA-based intermediaries that serve as proxies for direct genetic trait identification. These molecular intermediaries enable breeders to select for desirable traits based on genetic markers rather than waiting for phenotypic expression, improving selection precision and enabling earlier selection decisions in the breeding process
2Productivity
If molecular breeding techniques are used to improve efficiency and screening capability, then the breeding gain increases, but the process is still constrained by the number and locations of recombination events
Solution Approach 1:
The patent employs preliminary molecular screening and marker-assisted selection before final breeding decisions are made. By identifying desirable genetic traits at early stages using DNA markers, the method prepares and pre-selects candidate plants, improving breeding efficiency and reducing dependence on limited recombination events in later generations
Solution Approach 2:
The patent divides the genome into selectable segments using molecular markers that identify specific genetic regions of interest. This segmentation allows breeders to independently track and select for multiple desirable traits located at different genomic positions, effectively overcoming the constraints imposed by limited recombination events through marker-guided assembly of desired genetic segments
3Manufacturing precision
If custom endonucleases are introduced to modify specific loci, then the genetic modification precision is improved, but the device complexity and operational complexity increase
Solution Approach 1:
The patent utilizes the plant cell's own molecular machinery to execute the genetic modification. Custom endonucleases are introduced to create targeted DNA cuts, and the cell's endogenous repair mechanisms (homologous recombination or non-homologous end joining) perform the actual modification. This self-service approach achieves high precision genetic modification while reducing the complexity of external intervention systems
Solution Approach 2:
The patent employs custom endonucleases as molecular intermediaries that guide the modification process to specific genomic loci. These engineered proteins serve as precise delivery vehicles for creating targeted DNA breaks, while the cell's natural repair pathways complete the modification. This intermediary approach concentrates the complexity in a single, well-defined molecular tool rather than requiring complex external modification systems
4Stability of the object's composition
If recombination or meiosis is inhibited to maintain genetic modifications, then the stability of modified traits is improved, but the breeding process loses genetic diversity
Solution Approach 1:
The patent performs genetic modifications using custom endonucleases and targeted recombination before inhibiting further recombination or meiosis. By establishing the desired genetic composition in advance through directed molecular breeding, the method locks in the modifications while minimizing the need for subsequent recombination events, thereby maintaining stability without completely eliminating diversity-generating processes
Data Source
AI summary
The invention provides novel uses of sequence-specific or sequence-directed endonucleases for molecular plant breeding. The invention also provides novel plant transformation vectors and expression cassettes, which include novel combinations of an endonuclease with plant expression and transformation elements. Plants and derivatives thereof produced by such methods are also provided.