Aptamer-Mediated Non-Viral CRISPR Delivery
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Solution Overview
Problem
Current gene editing delivery systems, such as those using lentiviral vectors, face challenges like low transfection efficiencies and reduced cell viability, limiting the effective introduction of payloads like CRISPR-RNPs and gRNAs into cells.
Innovation Solution
Aptamer-mediated delivery systems comprising aptamers with specific binding domains and hybridization sequences are used to internalize payloads, including CRISPR components, by binding to cell surface molecules and facilitating internalization, potentially enhancing delivery efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If lentiviral vectors are used for payload delivery, then delivery capability is achieved, but transfection efficiency is low and cell viability is reduced
Solution Approach 1:
The patent uses aptamers as intermediary molecules that mediate between the payload (gene editing machinery) and the cell surface. The aptamers bind to cell surface receptors and facilitate payload delivery without requiring viral infection mechanisms, thereby improving transfection efficiency while maintaining delivery capability
Solution Approach 2:
The patent replaces the biological viral infection mechanism (lentiviral vectors) with a non-viral aptamer-mediated delivery system. This substitution eliminates the harmful effects of viral transduction while maintaining the ability to deliver payloads into cells, thus improving both transfection efficiency and cell viability
2Reliability
If lentiviral vectors are used for payload delivery, then delivery capability is achieved, but cell viability is reduced
Solution Approach 1:
Aptamers serve as safe intermediary molecules that enable payload delivery without the cytotoxic effects associated with lentiviral vectors. The aptamers bind to cell surface receptors and facilitate controlled payload entry, eliminating harmful viral components while preserving cell viability
Solution Approach 2:
The patent employs transient aptamer molecules that perform their delivery function and are then degraded or cleared from the system. These disposable aptamers eliminate the persistent harmful effects of integrated viral vectors while maintaining effective payload delivery, thus preserving long-term cell viability
3Productivity
If aptamers are used for payload delivery, then transfection efficiency is improved, but delivery mechanism complexity increases
Solution Approach 1:
The patent divides the delivery system into distinct functional modules: aptamers for cell surface binding, hybridization sequences for payload attachment, and payload components for gene editing. This segmentation allows each component to be optimized independently while simplifying the overall assembly and understanding of the delivery mechanism
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 potentially improves the efficiency of payload delivery into cells, reducing the drawbacks of existing systems by utilizing aptamers to target and internalize gene editing machinery, such as CRISPR components, with higher specificity and viability.
Implementation Method 1
the hybridization sequence of the first aptamer is configured to hybridize to the hybridization sequence of the second aptamer
Implementation Method 2
a cell surface binding domain... binding of the first aptamer to a cell surface molecule
Data Source
AI summary
Provided herein are compositions, systems, methods, and kits for internalizing a payload into a cell via aptamer-mediated deliver. Payloads can comprise molecules capable of hybridizing to nucleic acid sequences directly or through intermediaries (e.g. payload handles). More specifically, payloads can comprise gene editing machinery. A non-limiting example of gene editing machinery can comprise Clustered Regularly Interspaced Short Palindromic Repeats (“CRISPR”) RNP complexes or the subcomponents to those complexes (e.g. gRNA molecules and endonucleases).