Cas13 Viral RNA Co-Replication for Load-Responsive Antiviral Therapy
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current drug development for viral diseases, such as those caused by RNA viruses like SARS-CoV-2, is slow and ineffective due to the need for repurposing existing drugs and the potential for viral mutations to evade therapeutic agents, while existing CRISPR/Cas systems face challenges in efficiently targeting and inhibiting viral RNA.
Innovation Solution
A novel CRISPR/Cas13 system that includes nucleotide sequences encoding Cas13 protein and gRNA, flanked by viral 5′ and/or 3′ UTRs, which utilizes the viral replicase recognition sequence for self-amplification within infected cells, enhancing therapeutic efficiency by co-replicating with the virus and degrading viral mRNA.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional CRISPR/Cas systems are used to target viral RNA, then viral RNA can be degraded, but the system lacks efficient self-amplification and therapeutic efficacy is insufficient
Solution Approach 1:
The CRISPR/Cas13 system is designed to co-replicate with the virus using viral replicase recognition sequences in the 5' and 3' UTRs. The system serves itself by utilizing the viral replication machinery to amplify the CRISPR components, achieving automatic dose adjustment according to viral load without requiring external intervention or high initial doses
Solution Approach 2:
The system incorporates viral replicase recognition sequences that enable feedback-driven amplification. The viral replicase recognizes and binds to the UTR sequences, creating a feedback loop where viral replication automatically drives CRISPR system amplification, ensuring the system scales with viral load and maintains effective therapeutic concentrations
2Loss of time
If existing drugs are repurposed for viral diseases, then development time is reduced, but viral mutations can evade therapeutic agents
Solution Approach 1:
The invention extracts and targets the viral genetic information (RNA) directly rather than relying on protein targets that are susceptible to mutation. By using CRISPR/Cas13 to degrade viral RNA at the genome level, the system bypasses the limitation of protein-based therapies that can be evaded by viral mutations
Solution Approach 2:
The system dynamically adapts to viral variations through its self-amplifying mechanism. The CRISPR components are continuously replicated and amplified within the infected cell alongside the viral genome, ensuring that the therapeutic activity scales with the viral population and maintains effectiveness despite viral mutations
3Speed
If high initial doses of therapeutic agents are used, then rapid viral elimination can be achieved, but the cost and complexity increase
Solution Approach 1:
The system eliminates the need for high initial doses by designing a self-amplifying CRISPR/Cas13 system that utilizes viral replication to generate therapeutic components. The system automatically adjusts its own concentration based on viral load, achieving rapid viral elimination without requiring complex dose administration protocols or high initial therapeutic loads
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 system achieves an 85% reduction in viral load in cell culture experiments and demonstrates a wave-like correlation with viral load, allowing for rapid and effective elimination of viruses like SARS-CoV-2 with lower initial doses, particularly in severe disease progressions.
Implementation Method 1
at least one gRNA or at least one nucleotide sequence encoding said at least one gRNA capable of hybridizing with one or more viral target RNA molecules
Implementation Method 2
The novel system utilizes the RNase activity of the so-called Cas13 protein. In particular, a viral mRNA that has been introduced into or replicated in the cell is specifically degraded by the novel clustered, regularly interspaced, short palindromic repeats (CRISPR) system comprising at least one nucleotide sequence encoding at least one CRISPR-associated protein 13 (Cas13)
Implementation Method 3
a viral replicase recognition sequence is comprised in at least any one of said 5′ UTR or in the nucleotide sequence encoding said 5′ UTR, or said 3′ UTR or in the nucleotide sequence encoding said viral 3′ UTR
Data Source
AI summary
The present invention relates to a novel CRISPR system comprising i) at least one nucleotide sequence encoding at least one Cas13 protein; and ii) at least one gRNA or at least one nucleotide sequence encoding said at least one gRNA capable of hybridizing with one or more viral target RNA molecules, wherein said system comprises a viral 5′ UTR or a nucleotide sequence encoding said 5′ UTR and/or a viral 3′ UTR or a nucleotide sequence encoding said viral 3′ UTR, wherein a viral replicase recognition sequence is comprised in at least any one of said 5′ UTR or in the nucleotide sequence encoding said 5′ UTR, or said 3′ UTR or in the nucleotide sequence encoding said 3′ UTR, and wherein said system does not comprise a nucleotide sequence encoding a viral replicase and wherein said viral replicase recognition sequence is from the same RNA virus as the one or more viral target RNA molecules. The present invention also relates to a delivery system comprising the novel system and a composition comprising the novel system or the delivery system. The present invention further relates to the medical use of the novel system or in particular to the system for use in a method of preventing or treating a viral disease in a subject. Additionally, the present invention also relates to a kit comprising the novel system and to a method of producing the novel system.