Dried DNA Solid Support for Plant Protoplast Transformation
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Solution Overview
Problem
Current methods for high-throughput analysis of plant protoplasts are limited by the need for liquid culture, which leads to cell aggregation, movement, and heterogeneity, making microscopy-based analysis challenging and unsuitable for high-content screening.
Innovation Solution
A method involving the use of dried DNA immobilized on a solid support for transformation of plant protoplasts, combined with robotic handling and automated microscopy for high-throughput single cell analysis, allowing for reproducible transformation efficiencies and immobilization of cells in a monolayer for precise imaging.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If liquid culture is used for plant protoplasts, then cell growth and proliferation are maintained, but cell aggregation, movement, and heterogeneity occur making microscopy-based analysis challenging
Solution Approach 1:
The patent uses a thin film solid support surface to immobilize plant protoplasts, creating a stable monolayer that prevents cell aggregation and movement while maintaining cell viability and growth capabilities. This thin film approach allows precise microscopy-based analysis without the heterogeneity problems of liquid culture.
Solution Approach 2:
The patent changes the physical state of the culture medium from liquid to solid support surface, transforming the culture conditions while maintaining cell growth. This parameter change enables both cell proliferation and precise microscopy analysis by providing a stable, immobilized cell layer.
2Ease of manufacture
If DNA is dissolved in liquid for transformation, then transformation can be performed, but transformation efficiency is variable and not reproducible
Solution Approach 1:
The patent changes the physical state of DNA from dissolved liquid form to dried solid form immobilized on a solid support. This parameter change provides stable, controlled DNA delivery and significantly improves the reproducibility and reliability of transformation efficiency while maintaining ease of transformation process.
Solution Approach 2:
The patent replaces the liquid-based DNA delivery system with a solid-based system where DNA is immobilized on a solid support. This substitution provides more controlled and reproducible DNA transfer to protoplasts, eliminating the variability associated with liquid DNA solutions.
3Ease of operation
If manual handling methods are used for protoplast analysis, then flexibility is maintained, but high-throughput analysis and automation are limited
Solution Approach 1:
The patent creates a universal solid support system that can be used for both manual handling operations and automated high-throughput analysis. The standardized solid support with immobilized DNA and cells enables flexible manual operations while also being compatible with robotic handling and automated microscopy systems, achieving both ease of operation and high productivity.
Solution Approach 2:
The patent uses solid support surfaces that can be replicated and used in multi-well formats, enabling high-throughput analysis. The standardized solid support structure allows for automated handling and processing of multiple samples simultaneously while maintaining the flexibility needed for various experimental operations.
4Reliability
If cells are cultured in liquid medium, then cell viability is maintained, but cell movement and aggregation prevent stable imaging conditions
Solution Approach 1:
The patent uses a thin film solid support to immobilize cells, preventing movement and aggregation while maintaining cell viability. This approach provides stable imaging conditions by fixing cells in a monolayer configuration, eliminating the focus and position instability problems of liquid culture.
Solution Approach 2:
The patent changes the physical state of the culture environment from liquid to solid support, which stabilizes cell position for imaging while maintaining cell viability through appropriate solid support conditions. This parameter change resolves the contradiction between cell viability and imaging stability.
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 efficient, reproducible, and automated high-throughput analysis of plant protoplasts, overcoming the limitations of liquid culture by providing stable and controlled conditions for microscopy, thereby improving data quality and transformation efficiency.
Implementation Method 1
providing a solid support having immobilized thereto the polynucleotide in dry state
Implementation Method 2
contacting the plant protoplasts with the polynucleotide on the solid support so as to obtain transformed plant protoplast cells
Data Source
AI summary
The present invention relates to a method for introducing a polynucleotide into non-adhesively growing plant cells, comprising the following steps: providing a solid support having immobilized thereto the polynucleotide in dry state; contacting the plant cells with the polynucleotide on the solid support so as to obtain transformed plant cells; and optionally washing the plant cells.