SARS-CoV-2 Safe Replicon System for Drug Screening
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Solution Overview
Problem
Current SARS-COV-2 safe replicon systems, particularly those based on Bacterial Artificial Chromosomes (BAC), are unstable and require high-level biosafety laboratories for operation, limiting drug development and screening due to their large molecular weight and operational complexity.
Innovation Solution
A novel SARS-COV-2 safe replicon system comprising nucleic acid sequences encoding non-structural proteins, 5' and 3' UTRs, and transcription regulatory regions, integrated into expression vectors for use in low-level biosafety laboratories, allowing for the simulation of virus replication and drug screening.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If BAC plasmid is used to construct safe replicon system, then the system can simulate coronavirus replication, but the system becomes unstable and difficult to operate due to large molecular weight
Solution Approach 1:
The patent divides the large BAC plasmid into multiple smaller expression vectors (e.g., pcDNA3.1-based vectors). Each vector contains specific viral genes (such as ORF1ab, S, E, M, N genes) rather than the entire genome, making them easier to manipulate and more stable while collectively achieving the replication simulation function
Solution Approach 2:
The patent creates simplified copies of the viral replication system using only essential genes expressed in eukaryotic cells, rather than using the complete viral genome. This copy-based approach maintains replication simulation capability while eliminating the complexity and instability of the full BAC system
2Reliability
If BAC plasmid is used for safe replicon system, then virus replication can be simulated, but operation becomes time-consuming and laborious
Solution Approach 1:
By segmenting the replicon system into modular expression vectors, the patent enables independent manipulation of each vector without handling the entire large BAC plasmid. This segmentation significantly reduces the time and labor required for cloning, transformation, and maintenance operations
Solution Approach 2:
The patent changes the fundamental parameter of plasmid size from large (BAC) to small (standard expression vectors), which directly impacts operational efficiency. Smaller vectors are faster to transform, easier to propagate, and require less time for routine manipulations while maintaining the essential replication simulation function
3Measurement precision
If wild-type SARS-COV-2 is used for drug screening, then accurate drug response can be evaluated, but high-level biosafety laboratories are required which limits drug development
Solution Approach 1:
The patent creates a safe replicon system that copies only the essential replication machinery of SARS-COV-2 in the form of expression vectors. This copy allows accurate evaluation of antiviral drug effects on viral replication without requiring the complete, infectious wild-type virus, thereby enabling drug screening in lower biosafety level laboratories
Solution Approach 2:
The safe replicon system acts as an intermediary between the wild-type virus and the drug screening process. It provides a safe, controllable platform that mediates the evaluation of drug effects on viral replication, eliminating the need for direct handling of high-risk wild-type virus while maintaining measurement accuracy
Data Source
AI summary
The present disclosure discloses a novel coronavirus SARS-COV-2 safe replicon system and use thereof in screening an anti-SARS-COV-2 drug. The safe replicon system specifically comprises a nucleic acid sequence encoding a novel coronavirus SARS-COV-2 non-structural protein; and nucleic acid sequences of 5′ UTR and 3′ UTR of a novel coronavirus SARS-COV-2, a transcription regulatory region on which the novel coronavirus SARS-COV-2 non-structural protein can act, and a reporter gene. With the SARS-COV-2 safe replicon system, high-throughput screening of anti-SARS-COV-2 drugs and pharmacologic verification of drugs can be carried out independent of a biosafety level 3 laboratory, and the operation is simple and convenient.