Cell Molecule Introduction via Lysosomal Inhibition
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Solution Overview
Problem
Current methods for introducing selected molecules into cells are inefficient due to degradation within the cell, leading to a substantial decrease in the molecule's ability to exert its function.
Innovation Solution
A method involving an inhibitor treatment step to block endocytosis- or autophagy-mediated lysosomal degradation pathways, combined with the introduction of a selected molecule via endocytosis or plasma irradiation, using low-molecular-weight compounds like nocodazole, chloroquine, or LY-294002 to stabilize the molecule within the cell.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If a selected molecule is introduced into a cell using conventional methods (virus, electroporation, lipofection, or plasma), then the molecule can be delivered into the cell, but the molecule is degraded intracellularly by lysosomal degradation pathways, resulting in decreased functional efficiency
Solution Approach 1:
The invention applies preliminary anti-action by treating cells with lysosomal degradation pathway inhibitors (such as chloroquine, bafilomycin A1, or E64d) before introducing the selected molecule. This pre-treatment blocks the lysosomal degradation machinery in advance, preventing the molecule from being degraded after introduction, thereby resolving the contradiction between molecule stability and introduction efficiency
Solution Approach 2:
The invention uses preliminary action by preparing cells with inhibited lysosomal degradation pathways before molecule introduction. The inhibitor treatment is performed in advance to alter the cellular state, creating a protective environment that prevents subsequent molecule degradation while maintaining efficient introduction capabilities
2Reliability
If the concentration of inhibitor is increased to further block degradation pathways, then molecule stability improves, but cytotoxic effects increase and cell viability decreases
Solution Approach 1:
The invention applies parameter changes by optimizing the inhibitor concentration to a specific range (0.01-10 μM for chloroquine, 0.001-1 μM for bafilomycin A1, or 0.01-1 μM for E64d) and treatment time (0.5-24 hours). This precise parameter control achieves sufficient lysosomal pathway inhibition for molecule stability while maintaining cell viability and minimizing cytotoxic effects
Solution Approach 2:
The invention uses partial action by applying a moderate level of inhibition rather than complete blockade. The inhibitor concentration is optimized to achieve sufficient protection against molecule degradation without overwhelming the cellular systems, thereby avoiding excessive cytotoxic effects and maintaining cell health
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 significantly increases the efficiency of introducing selected molecules into target cells, enhancing their stability and functionality, while minimizing cytotoxic effects and maintaining cell viability.
Implementation Method 1
an introduction step of contacting an introduction liquid containing a selected molecule with the target cell or the target tissue to introduce the selected molecule into the target cell or the target tissue
Implementation Method 2
an inhibitor treatment step of allowing a target cell or a target tissue to internalizing an inhibitor of an endocytosis- or autophagy-mediated lysosomal degradation pathway
Implementation Method 3
in the introduction step, the selected molecule is introduced by irradiating the target cell or the target tissue with plasma
Data Source
AI summary
Provided is a method for introducing a selected molecule, including: an inhibitor treatment step of allowing a target cell or a target tissue to internalizing an inhibitor of vesicular transport or lysosomal membrane fusion; and an introduction step of contacting an introduction liquid containing a selected molecule with the target cell or the target tissue to introduce the selected molecule into the target cell or the target tissue.