Self-Assembling Polypeptide Nanoparticles for Nucleic Acid Delivery
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Solution Overview
Problem
Current transfection tools are ineffective for delivering nucleic acids into hard-to-transfect cells, such as primary neuronal cultures, often requiring harsh or toxic methods.
Innovation Solution
Engineered polypeptides that self-assemble into nanoparticles upon binding to nucleic acids, facilitating efficient cellular uptake and delivery by incorporating a nucleic acid binding domain and an assembly domain that forms secondary structure upon complex formation, enabling targeted cell internalization and endosome escape.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional transfection tools are used, then transfection of easy-to-transfect cells is achieved, but transfection of hard-to-transfect cells (e.g., primary neuronal cultures) is ineffective
Solution Approach 1:
The invention changes the physical and chemical parameters of the transfection system by using self-assembling polypeptide nanoparticles with specific assembly domains and nucleic acid binding domains. These nanoparticles form complexes with nucleic acids that can be internalized by a broad range of cell types including hard-to-transfect primary neuronal cultures, achieving reliable transfection across diverse cell types without requiring cell-type-specific optimization
Solution Approach 2:
The invention creates composite nanoparticle structures consisting of polypeptides with multiple functional domains (assembly domain + nucleic acid binding domain) that self-assemble into nanoparticles. These composite structures combine the benefits of self-assembly for efficient cellular uptake with specific nucleic acid binding capabilities, enabling effective transfection of previously difficult-to-transfect cell types
2Reliability
If harsh or toxic methods are used to achieve transfection of hard-to-transfect cells, then transfection efficiency improves, but cell toxicity increases
Solution Approach 1:
The polypeptides perform self-service by automatically self-assembling into nanoparticles upon binding to nucleic acids, without requiring external assembly machinery or harsh chemical treatments. This self-assembly process occurs under physiological conditions and generates nanoparticles that are efficiently internalized by cells, achieving high transfection efficiency without the need for toxic external forces
Solution Approach 2:
The invention replaces harsh mechanical or chemical transfection methods (such as sonication, electroporation, or chemical detergents) with a biochemical self-assembly mechanism. The polypeptides use their intrinsic assembly domain to spontaneously form nanoparticles that facilitate nucleic acid delivery, substituting physical/chemical force-based methods with a gentle, biology-based self-organization process that minimizes cell stress and toxicity
3Productivity
If polypeptides self-assemble into nanoparticles upon binding to nucleic acids, then cellular internalization efficiency increases, but polypeptide structure complexity increases
Solution Approach 1:
The polypeptide is segmented into distinct functional domains: an assembly domain that mediates self-assembly into nanoparticles and a nucleic acid binding domain (NABD) that binds to the cargo. This segmentation allows each domain to perform its specific function independently, with the assembly domain driving nanoparticle formation and the NABD ensuring nucleic acid complexation, thereby achieving high internalization efficiency through modular design
Solution Approach 2:
The assembly domain serves multiple functions: it mediates self-assembly of polypeptides into nanoparticles, provides structural organization for the nanoparticle, and facilitates efficient cellular internalization. This multi-functional assembly domain reduces the need for additional separate components, achieving high productivity through a streamlined, multi-functional polypeptide design
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
The polypeptide/nucleic acid complexes are internalized by cells with high efficiency and low toxicity, effectively triggering biological effects like gene knockdown in neuronal cells, surpassing commercial controls in internalization and gene expression while minimizing toxicity.
Implementation Method 1
the assembly domain forms secondary structure with assembly domains of adjacent polypeptide in the presence of complex formation between the NABD and nucleic acids. In some embodiments, the secondary structure comprises beta-sheet interactions.
Implementation Method 2
the assembly domain facilitates self-assembly of the polypeptide/nucleic acid complex into nanoparticles
Implementation Method 3
the assembly domain forms supramolecular interactions with assembly domains of adjacent polypeptide in the presence of complex formation between the NABD and nucleic acids. In some embodiments, the supramolecular interactions comprise hydrogen bonds.
Data Source
AI summary
Provided herein are polypeptides that self-assemble into nanoparticles upon binding to nucleic acids. The nanoparticles find use, for example in the delivery of the nucleic acids into cells.