Repressible Operon Viral Packaging Vectors for Higher Particle Yield
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Solution Overview
Problem
Current viral vector systems face challenges in effectively packaging exogenous genes due to unpredictable expression effects that hinder viral particle production, leading to reduced yields and inefficiencies.
Innovation Solution
A vector system with inverted terminal repeats and a repressible operon is used to regulate exogenous gene expression, allowing packaging in the absence of a repressor to inhibit gene expression, thereby enhancing viral particle production efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If exogenous gene is driven by strong promoter during viral packaging, then gene expression level is high, but viral particle yield is reduced due to unpredictable negative effects on packaging processes
Solution Approach 1:
The patent applies preliminary action by introducing a repressible operon system before viral packaging to prevent unwanted gene expression. The repressor protein is expressed during packaging to silence exogenous gene expression, and can be removed afterward to enable gene expression in target cells. This preliminary control mechanism resolves the contradiction by preventing packaging interference while preserving therapeutic functionality.
2Productivity
If exogenous gene expression is inhibited during packaging using repressible operon, then viral particle yield is improved, but gene expression capability in target cells may be compromised
Solution Approach 1:
The patent applies dynamics by creating a dynamic, controllable gene expression system. The repressible operon allows gene expression to be switched on or off based on the presence or absence of repressor. During packaging, repressor is present to inhibit expression and maximize viral particle production. After packaging, repressor can be removed to enable gene expression in target cells. This dynamic control resolves the contradiction by allowing both high viral yield and effective gene delivery.
3Device complexity
If conventional viral packaging is used without gene expression regulation, then packaging process is simple, but exogenous gene has unpredictable negative effects on viral particle production
Solution Approach 1:
The patent applies the intermediary principle by introducing a repressible operon system as a mediator between the exogenous gene and the packaging process. This intermediary mechanism allows controlled gene expression during packaging, preventing negative effects on viral particle production. The repressor protein acts as the intermediary that can be added or removed to control expression timing, resolving the contradiction between simplicity and productivity.
Data Source
AI summary
The present application relates to the technical field of biology, and specifically, to an exogenous gene controllable expression virus packaging vector and a packaging method. The virus packaging vector has two ITR fragments and a gene expression cassette inserted between the two ITR fragments. The gene expression cassette comprises a promoter, a repressor operator, and an optional target gene that are connected in sequence. When there is a repressor, the repressor operator can repress the expression of a downstream target gene thereof.