β-Helical Protein Conjugates for Endocytosis-Free Gene Editing Delivery
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for delivering gene-editing molecules across cellular membranes, particularly for gene therapy and genome editing, face inefficiencies and challenges, including endocytosis-dependent mechanisms that can trap or degrade the molecules, and limitations of viral and non-viral delivery systems, especially for tissues like lung epithelium and skeletal muscle.
Innovation Solution
A recombinant β helical protein, ranging from 5 to 25 nm in length and 1 to 5 nm in width, is linked to genome-editing molecules or plasmids to form a genome-editing complex, utilizing non-covalent interactions and a signal sequence for targeted cell delivery, enhancing membrane penetration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If endocytosis-dependent mechanisms are used for delivery, then molecules can enter cells, but the molecules get trapped or degraded in endosomes/lysosomes
Solution Approach 1:
The delivery system is segmented into two functional components: a cell-penetrating peptide (CPP) for membrane translocation and a functional molecule (gene-editing molecule, drug, or protein) for therapeutic action. This segmentation allows the CPP to handle membrane penetration while the functional molecule remains protected and active, avoiding degradation in endosomes/lysosomes.
Solution Approach 2:
The cell-penetrating peptide acts as an intermediary that mediates the transport of functional molecules across the cell membrane. The CPP conjugate serves as a carrier that facilitates direct cytoplasmic delivery, bypassing the endocytic pathway that would otherwise trap or degrade the functional molecule.
2Productivity
If viral delivery systems are used, then gene delivery efficiency is improved, but safety concerns and immunogenicity arise
Solution Approach 1:
The patent employs non-viral, disposable delivery vectors (CPP conjugates) that can be synthesized chemically or recombinantly. These short-lived, non-integrating delivery systems achieve transient gene delivery without the long-term safety concerns and immunogenicity associated with viral vectors, allowing repeated administrations if needed.
Solution Approach 2:
The delivery system transitions from viral to non-viral parameters, changing the fundamental nature of the vector. By using recombinant proteins or chemically synthesized peptides as delivery vehicles instead of viral particles, the system maintains delivery efficiency while eliminating viral-associated safety risks and immunogenicity.
3Object-affected harmful factors
If non-viral delivery systems are used, then safety is improved, but delivery efficiency to certain tissues (lung epithelium, skeletal muscle) is reduced
Solution Approach 1:
The cell-penetrating peptide component is specifically selected or engineered to have local quality properties optimized for penetrating certain tissue types. Different CPPs can be chosen or designed with specific characteristics (charge, hydrophobicity, structure) that enhance delivery to particular tissues like lung epithelium or skeletal muscle while maintaining the safety advantages of non-viral systems.
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 β helical protein-conjugate efficiently transfers genome-editing molecules into various cell types, including eukaryotic and prokaryotic cells, enabling effective gene editing and gene therapy by avoiding endocytosis and improving delivery efficiency.
Implementation Method 1
A recombinant β helical protein, ranging from 5 to 25 nm in length and 1 to 5 nm in width, is linked to genome-editing molecules or plasmids to form a genome-editing complex, utilizing non-covalent interactions
Data Source
AI summary
A genome-editing complex for modifying a target polynucleotide comprising a recombinant β helical protein linked to either one or more molecules of a genome-editing system or a plasmid encoding for one or more molecules of a genome-editing system, wherein the β helical protein length is in the range of from 5 nm to 25 nm, and width is in the range of from 1 nm to 5 nm.