Laser-aided mRNA Transfection for Cell Phenotype Conversion
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Solution Overview
Problem
Current methods for effecting phenotypic change in cells are complex and difficult to control, particularly in re-differentiating induced pluripotent stem cells into desired cell types, and there is a need for more efficient techniques to achieve phenotypic conversion in cells.
Innovation Solution
A method involving the transfection of phenotype-converting nucleic acid, such as mRNA transcriptome, from a donor cell into a recipient cell using laser-aided poration, allowing for precise control of nucleic acid introduction and expression, enabling changes in gene expression, protein expression, immunological markers, morphology, physiology, and membrane lipid composition.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If nuclear transplantation or transfection of transcription factors is used to achieve phenotypic change, then cellular phenotype can be converted, but the process becomes complex and difficult to control
Solution Approach 1:
The patent extracts and transfers only the essential phenotype-determining components (mRNA transcriptome, proteins, or small molecules) from donor cells to recipient cells, rather than transferring entire nuclei or using complex multi-factor transfection systems. This extraction approach simplifies the process while maintaining phenotype conversion capability.
Solution Approach 2:
The patent creates copies of phenotype-determining molecules (mRNA, proteins) from donor cells and introduces these copies into recipient cells. This copying approach allows precise control over the transferred material and simplifies the overall process compared to nuclear transplantation.
2Adaptability or versatility
If induced pluripotent stem cells are re-differentiated into desired cell types, then phenotypic change is achieved, but the process adds a layer of complexity that is difficult to control
Solution Approach 1:
The patent performs preliminary action by directly introducing phenotype-determining molecules into recipient cells to achieve the desired phenotypic state, bypassing the multi-step re-differentiation process required by traditional iPS cell methods. This direct approach reduces complexity while achieving the same cell type conversion outcome.
3Stability of the object's composition
If phenotype-converting nucleic acid is transfected into recipient cells, then stable phenotype conversion is achieved, but control over the transfection process is needed
Solution Approach 1:
The patent replaces mechanical transfection methods with optical manipulation (optical tweezers) to introduce phenotype-converting nucleic acid into recipient cells. This substitution provides precise spatial and temporal control over the transfection process while maintaining stable phenotype conversion, improving ease of operation compared to traditional mechanical methods.
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 method achieves stable phenotype conversion in recipient cells, with up to 35% of cells undergoing phenotype conversion, and the changes can persist for extended periods, including the lifetime of the cell, allowing for precise control of cellular phenotype conversion.
Implementation Method 1
transfection of phenotype-converting nucleic acid, such as mRNA transcriptome, from a donor cell into a recipient cell using laser-aided poration
Data Source
AI summary
The present invention includes methods for effecting phenotype conversion in a cell by transfecting the cell with phenotype-converting nucleic acid. Expression of the nucleic acids results in a phenotype conversion in the transfected cell. Preferably the phenotype-converting nucleic acid is a transcriptome, and more preferably an mRNA transcriptome.