Augmented sgRNA Mobile Delivery for Plant Genome Editing
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Solution Overview
Problem
Current methods for genome editing in plants face challenges due to inefficient delivery of gene editing reagents to somatic and germline cells, leading to low frequencies of heritable genetic modifications, particularly in plants where regeneration protocols are unavailable or genotype-dependent, and are time-consuming.
Innovation Solution
The use of augmented short guide RNAs (sgRNAs) that are modified to be mobile, allowing them to move from cell to cell and into the meristem, combined with viral vectors for delivery, enabling high-frequency germline editing by interacting with RNA-guided gene editing reagents like Cas9 for specific genomic modifications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional transformation methods (Agrobacterium-mediated T-DNA delivery or particle bombardment) are used to deliver gene editing reagents to plant cells, then editing can occur in recipient somatic cells, but the process requires time-consuming regeneration through callus formation and hormone treatment, taking several months to a year to generate edited plants
Solution Approach 1:
The patent extracts and utilizes endogenous transposable elements (TEs) from the plant genome itself as delivery vehicles for gene editing reagents. By hijacking the plant's own mobile genetic elements, the system eliminates the need for external transformation methods and lengthy regeneration protocols, directly delivering CRISPR components to germ cells and producing edited plants in a single generation without callus formation or hormone treatment
Solution Approach 2:
The patent employs the plant's own biological systems - specifically endogenous transposable elements and the plant's natural transformation capabilities - to deliver gene editing reagents. This self-service approach leverages the plant's intrinsic mechanisms for genetic movement and transformation, eliminating dependence on external delivery systems and complex tissue culture regeneration processes
2Ease of operation
If virus vectors are used to deliver gene editing reagents to plants, then delivery can be achieved, but the frequency of germline mutations recovered from resulting plants is extremely low, making the approach too inefficient for creating genetic variation
Solution Approach 1:
The patent fundamentally changes the delivery parameter from viral vectors to endogenous transposable elements. By switching to TEs that naturally integrate into the germ line and can be vertically transmitted, the system achieves high frequencies of heritable mutations. The TE-based delivery system exploits the plant's own genetic mobility mechanisms, resulting in significantly higher recovery rates of germline edits compared to viral vector approaches
3Reliability
If gene editing reagents are delivered to somatic plant cells via transformation, then editing can occur in recipient cells, but for many plant species protocols for regenerating plants from somatic cells are not available and success is genotype dependent, making the process technically challenging
Solution Approach 1:
The patent creates a universal gene editing delivery system based on endogenous transposable elements that can be applied across diverse plant species. Since TEs are naturally present in most plant genomes and have inherent mobility mechanisms, this approach eliminates the need for species-specific transformation protocols and regeneration procedures. The system works across different plant types without requiring genotype-dependent optimization, making gene editing broadly applicable and technically straightforward
Data Source
AI summary
Methods and materials for increasing somatic and germline genome editing are provided herein. For example, provided herein are methods and materials for using augmented sgRNAs to increase somatic and germline genome editing.