Virus-like Particles for Prime Editor Delivery
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Solution Overview
Problem
Current methods for delivering prime editors (PEs) in vivo face challenges such as prolonged expression leading to off-target editing and the risk of viral vector integration, which can promote oncogenesis or other adverse effects.
Innovation Solution
The development of virus-like particles (VLPs) engineered to package prime editors, associated guide RNAs, and other components, allowing for efficient prime editing while minimizing off-target effects by delivering proteins or ribonucleoproteins instead of DNA.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If viral vectors (AAV or LV) are used to deliver DNA encoding prime editors, then delivery efficiency to target tissues is improved, but prolonged expression occurs leading to off-target editing and risk of viral integration
Solution Approach 1:
The patent segments the delivery system by using virus-like particles (VLPs) that contain prime editor proteins and RNPs rather than viral DNA. This segmentation separates the delivery vehicle from the genetic material, allowing efficient cellular uptake while preventing prolonged expression and integration issues associated with viral DNA vectors.
Solution Approach 2:
The patent employs short-lived RNP complexes delivered via VLPs instead of persistent viral DNA. The RNP complexes are transient and degraded after completing their editing function, eliminating the prolonged expression problem that causes off-target editing. This disposable approach maintains high delivery efficiency while ensuring safety through limited duration of activity.
2Ease of operation
If viral vectors are used to deliver prime editor DNA, then target tissue delivery is achieved, but the DNA integrates into the genome promoting oncogenesis or adverse effects
Solution Approach 1:
The patent extracts the harmful integration capability from the delivery system by using VLPs containing proteins and RNPs instead of integrative viral DNA. This extraction removes the risk of genomic integration and oncogenesis while preserving the ability to deliver prime editors to target tissues through the VLP delivery mechanism.
Solution Approach 2:
The patent introduces VLPs as an intermediary delivery vehicle that bridges the gap between extracellular delivery and intracellular function without requiring genomic integration. The VLPs mediate delivery of prime editor proteins and RNPs into cells, eliminating the need for viral DNA integration while maintaining effective target tissue delivery.
3Adaptability or versatility
If transfection methods are used to deliver prime editors, then delivery to various cells is possible, but efficiency varies dramatically especially in primary cells
Solution Approach 1:
The patent creates a universal delivery system using VLPs that can deliver prime editors to multiple cell types including primary cells with high efficiency. The VLP structure provides multi-functional capability to transduce diverse cell types without the efficiency variation problems associated with conventional transfection methods, achieving both broad adaptability and high productivity.
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 use of VLPs enables safe and efficient delivery of prime editors, reducing the risk of off-target editing and adverse effects, while maintaining effective target editing capabilities.
Implementation Method 1
the components of the VLPs provided herein self-assemble at the cell membrane and bud out in accordance with the naturally occurring mechanism of budding (e.g., retroviral budding or the budding mechanism of other envelope viruses) in order to release from the cell fully-matured VLPs
Data Source
AI summary
The present disclosure provides virus-like particles (VLPs) for delivering prime editors, and systems comprising such prime editor (PE) VLPs. The present disclosure also provides polynucleotides encoding the PE-VLPs described herein, which may be useful for producing said PE-VLPs. Also provided herein are methods for editing the genome of a target cell by introducing the presently described PE-VLPs into the target cell. The present disclosure also provides fusion proteins that make up a component of the PE-VLPs described herein, as well as polynucleotides, vectors, cells, and kits.