Wheat Genome Editing via Ribonucleoprotein Integration
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Solution Overview
Problem
Traditional wheat breeding methods are inefficient and introduce random genetic or epigenetic variations, making precise modification of wheat genomes challenging, especially in recalcitrant regions and without the use of DNA or bacterially-mediated transformation.
Innovation Solution
The development of methods to make precise, non-random modifications in wheat genomes using ribonucleoproteins with sequence-specific nucleases and guide RNAs, allowing for modifications in specific alleles or genes without DNA transformation or selection, and without callus culture, ensuring high accuracy and minimal unwanted changes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If traditional wheat breeding methods are used, then wheat plants can be obtained, but random genetic or epigenetic variations are introduced and modification precision is poor
Solution Approach 1:
The patent segments the genome modification process into targeted steps using guide RNAs that direct nucleases to specific genomic loci. This allows precise modification of individual genes or regulatory elements without affecting other parts of the genome, thereby improving modification precision while maintaining genetic stability.
Solution Approach 2:
The patent uses guide RNAs as intermediaries to mediate between the nuclease enzyme and the target genomic sequence. The guide RNA provides sequence-specific recognition, enabling precise targeting of modifications while preventing random genetic variations, thus resolving the contradiction between precision and reliability.
2Productivity
If recalcitrant genome regions are modified using traditional methods, then modifications can be achieved, but efficiency is low and random variations increase
Solution Approach 1:
The patent replaces traditional mechanical breeding methods (crossing, selection) with a molecular-based system using ribonucleoproteins and guide RNAs. This substitution enables direct, precise editing of recalcitrant genome regions without relying on random recombination events, thereby improving both efficiency and accuracy of modifications.
3Reliability
If DNA transformation or bacterially-mediated transformation is used, then gene integration can be achieved, but unwanted genetic variation and epigenetic changes are introduced
Solution Approach 1:
The patent extracts and eliminates the need for DNA transformation and bacterial mediation by using ribonucleoprotein complexes that directly edit the genome in vivo. This removal of intermediate steps prevents the introduction of unwanted genetic variations and epigenetic changes while maintaining precise allele modification capability.
4Manufacturing precision
If callus culture is used for genome modification, then modified plants can be regenerated, but random genetic variations and epigenetic changes occur
Solution Approach 1:
The patent performs genome modification directly in the plant cell or tissue before regeneration, using ribonucleoprotein complexes that edit the genome in its native context. This preliminary action at the cellular level avoids the need for callus culture, thereby preventing random genetic variations and epigenetic changes while maintaining modification accuracy.
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 enables precise modification of wheat genomes with minimal random genetic or epigenetic changes, resulting in improved traits such as abiotic stress tolerance, disease resistance, and modified architecture, while maintaining over 99.9% identity to the reference genome.
Implementation Method 1
integration of a sequence encoded by a donor polynucleotide molecule at the site of at least one double-strand break in a genome
Data Source
AI summary
The disclosure relates to precise modifications in wheat plant genomes including the heterologous integration of nucleic acid sequences that include a regulatory element at a predetermined locus. Also provided are wheat cells, wheat plants, wheat seeds, and processed wheat products comprising the modified wheat genomes.