Microfluidic Delivery of Gene Editing Components
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Solution Overview
Problem
Current genome editing technologies face challenges in delivering gene editing components, such as protein and nucleic acid complexes, across cellular membranes, often requiring harmful viral vectors or electric fields, and existing methods are inefficient and prone to off-target effects.
Innovation Solution
A method involving mechanical cell disruption using a microfluidic system with cell-deforming constrictions to deliver protein-nucleic acid complexes, such as Cas9 and guide RNA, directly into cells, avoiding the need for viral vectors and minimizing off-target effects by complexing these components before delivery.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If viral vectors or electric fields are used to deliver gene editing components, then delivery capability is achieved, but harmful effects and safety issues arise
Solution Approach 1:
The patent extracts and eliminates the harmful viral vectors and electric fields from the delivery process. Instead of using these traditional methods, the invention employs a microfluidic device with a constriction that mechanically disrupts the cell membrane in a controlled manner, allowing direct delivery of protein-nucleic acid complexes without the harmful side effects of viral integration or electrical damage
Solution Approach 2:
The microfluidic constriction acts as an intermediary mechanism between the external environment and the cell interior. It provides a controlled mechanical disruption pathway that facilitates the entry of gene editing complexes without requiring harmful viral vectors or electric fields, thus mediating the delivery process safely
2Ease of manufacture
If gene editing components are delivered without complexing, then delivery simplicity is maintained, but off-target effects increase
Solution Approach 1:
The patent applies preliminary action by complexing the protein and nucleic acid components before delivery through the microfluidic constriction. This pre-complexing ensures that the gene editing components are stabilized and directed toward the cell interior, reducing off-target effects while maintaining relative simplicity in the overall delivery process
Solution Approach 2:
The invention changes the physical and chemical parameters of the gene editing components by forming protein-nucleic acid complexes. This complexing modifies the delivery characteristics and cellular uptake mechanisms, thereby reducing off-target effects while maintaining ease of delivery through the microfluidic system
3Manufacturing precision
If complexing is performed before delivery, then delivery precision is improved, but process complexity increases
Solution Approach 1:
The patent performs the complexing action as a preliminary step before the actual delivery process. By pre-complexing the protein and nucleic acid components in solution before they enter the microfluidic device, the invention achieves precise delivery without significantly increasing the overall process complexity, as the complexing occurs in a simple mixing chamber
4Ease of manufacture
If traditional delivery methods are used, then existing protocols are maintained, but efficiency and safety are compromised
Solution Approach 1:
The patent replaces the traditional biological or electrical mechanisms (viral vectors, electric fields) with a controlled mechanical system - the microfluidic constriction that physically disrupts the cell membrane. This substitution achieves higher efficiency and safety while maintaining ease of use through a standardized mechanical process that can be integrated into existing laboratory protocols
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 and specific gene editing by ensuring the complexes are fully functional upon entry into the cytosol, reducing exposure time and minimizing unwanted interactions, thereby improving the precision and safety of genome engineering applications.
Implementation Method 1
passing the cell through the constriction such that a pressure is applied to the cell causing perturbations of the cell large enough for said protein-nucleic acid complex to pass through
Data Source
AI summary
Gene editing can be performed by introducing gene-editing components into a cell by mechanical cell disruption. Related apparatus, systems, techniques, and articles are also described.