Mobile Genetic Element Integration for Non-Viral Immune Cell Engineering
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Solution Overview
Problem
Current methods for delivering large nucleic acid cargo into cells for therapeutic purposes face safety issues and inefficiencies, particularly with viral delivery mechanisms, and there is a need for stable, non-viral integration and expression of genetic material in cells, especially in immune cells for immunotherapy.
Innovation Solution
The use of a pharmaceutical composition comprising a mobile genetic element with a sequence encoding a polypeptide and an insert sequence that integrates into the genome of a cell, utilizing non-viral methods such as target-primed reverse transcription and endonuclease-mediated integration, allowing for stable integration of large nucleic acid sequences into immune cells like T cells, B cells, and macrophages.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If viral delivery mechanisms are used to deliver large nucleic acid cargo, then delivery efficiency is improved, but safety issues arise and immunogenicity increases
Solution Approach 1:
The patent uses a non-viral intermediary system consisting of mobile genetic elements (such as retrotransposons) and cellular enzymes (reverse transcriptase, integrase) to mediate the delivery and integration of large nucleic acid cargo. This intermediary approach replaces viral vectors with a safer, cellular-based mechanism that achieves efficient delivery without the immunogenicity and safety concerns of viral systems
Solution Approach 2:
The patent substitutes the mechanical/viral delivery system with a biochemical system utilizing cellular machinery. Instead of relying on viral particles to physically deliver cargo, the system uses mobile genetic elements that hijack endogenous cellular enzymes (reverse transcriptase, integrase) to perform integration, replacing a foreign mechanical system with the cell's own biochemical machinery
2Reliability
If repeated gene manipulation is performed on cells, then genetic modification is achieved, but cell health deteriorates and cell cycle alterations occur
Solution Approach 1:
The patent enables the cell to perform its own genetic modification by introducing mobile genetic elements that encode their own integration machinery. The cell's endogenous enzymes (reverse transcriptase, integrase) are recruited to perform the integration, making the system self-service and reducing the need for repeated external manipulation that would harm cell health
Solution Approach 2:
The patent introduces mobile genetic elements that carry the necessary genetic information and integration capability in advance. This preliminary action allows the cell to autonomously complete the genetic modification process without requiring multiple rounds of external intervention, thereby preserving cell health and avoiding cell cycle disruptions
3Object-affected harmful factors
If non-viral methods are used for genetic integration, then safety is improved, but integration efficiency decreases
Solution Approach 1:
The patent introduces mobile genetic elements as intermediaries that bridge the gap between safe non-viral delivery and efficient integration. These elements carry both the therapeutic cargo and the genetic instructions for their own integration, utilizing cellular machinery to achieve efficient integration without the safety compromises of viral vectors
Solution Approach 2:
The mobile genetic elements serve multiple functions simultaneously: they deliver the therapeutic cargo, encode the integration machinery, and utilize endogenous cellular enzymes for integration. This multi-functionality allows a single non-viral system to achieve both safety and integration efficiency that were previously thought to be mutually exclusive
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 safe, stable, and efficient integration and expression of large nucleic acid sequences, such as those encoding chimeric antigen receptors, in immune cells, facilitating effective immunotherapy without the drawbacks of viral vectors.
Implementation Method 1
via target-primed reverse transcription (TPRT)
Implementation Method 2
by cleavage of a DNA strand of a target site by an endonuclease encoded by the one or more polynucleic acids
Implementation Method 3
via reverse splicing of the insert sequence into a DNA target site of the genome
Data Source
AI summary
Methods and composition for modulating a target genome and stable integration of a transgene of interest into the genome of a cell are disclosed.