Non-epidermal Plant Cell Genome Editing via Particle Bombardment
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Solution Overview
Problem
Current genome editing technologies, such as CRISPR, face challenges in efficiently and precisely modifying genetic sequences in plant cells, particularly in non-epidermal cells capable of division and differentiation.
Innovation Solution
The method involves delivering a guide RNA (gRNA) to non-epidermal plant cells using various delivery techniques, such as electroporation or chemical agents, to alter specific target nucleotide sequences. This can include using CRISPR RNA (crRNA) or single guide RNA (sgRNA) in combination with an RNA-guided nuclease like Cas9 to introduce genetic alterations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If CRISPR technology is used to edit genes in plant cells, then precise genetic modification is achieved, but delivery efficiency to non-epidermal cells remains insufficient
Solution Approach 1:
The patent uses particle bombardment as an intermediary delivery mechanism to transport CRISPR components (Cas9 protein and guide RNA) into non-epidermal plant cells. Gold or tungsten particles coated with the CRISPR components are accelerated into the cells, serving as a mediator that overcomes the cell wall and membrane barriers, thereby improving delivery efficiency while maintaining genetic modification precision.
2Adaptability or versatility
If genome editing is performed in non-epidermal cells capable of division and differentiation, then enhanced traits can be propagated, but the complexity of delivering editing tools increases
Solution Approach 1:
The patent segments the CRISPR delivery system into separate components: the Cas9 protein, guide RNA, and particle bombardment mechanism. This segmentation allows each component to be optimized independently and simplifies the overall delivery process to non-epidermal cells, reducing system complexity while enabling effective genetic editing in cells capable of division and differentiation for trait propagation.
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 the precise modification of genetic sequences in plant cells, leading to the development of novel plant cells, plants, and seeds with enhanced traits, such as improved stress tolerance or altered metabolic pathways.
Implementation Method 1
delivering a guide RNA (gRNA) to a non-epidermal plant cell... wherein the gRNA has a nucleotide sequence designed to alter a target nucleotide sequence... In embodiments, the non-epidermal plant cell contains a nuclease, such as a Cas9 nuclease or other RNA-guided nuclease
Implementation Method 2
The methods include one or more delivery steps or treatments, including treatment with at least one chemical, enzymatic, or physical agent or use of techniques such as application of heat or cold, ultrasonication, centrifugation, and electroporation
Implementation Method 3
The methods include one or more delivery steps or treatments, including treatment with at least one chemical, enzymatic, or physical agent
Data Source
AI summary
Disclosed herein are compositions and methods for effecting alterations at a defined location in the genome of a non-epidermal plant cell. Further disclosed are methods for providing plants having a modified phenotype or a modified genome.