Application

Cycle 1: Timing of Intervention – Extending the Protection Window

• Acknowledge (The Problem): Our original system was designed as a post-exposure prophylactic, but we lacked clarity on the optimal timing for administration relative to pathogen exposure.
• Research (The Exchange): The hospital emphasized that "the entire flu season is suitable for vaccination" and that protection can be initiated at any time before infection occurs. This reflects a flexible, season-long prevention strategy rather than a narrow pre-exposure window.
• Effect (The Impact): We reconceptualized our platform as a "whole-season prophylactic", suitable for administration at any point during a respiratory virus season, not only after known exposure.
• Adjust (The Refinement): We are now testing the kinetics of CRISPRa activation and persistence in vitro to ensure our system remains effective even when administered mid-outbreak, mirroring the public health guidance that “it’s never too late to vaccinate.”

Cycle 2: Target Population – Broadening Applicability

• Acknowledge (The New Problem): Our initial target population was vaguely defined as “high-risk,” without clear stratification by age or comorbidity, just individuals who were classified as immunocompromised.
• Research (The Exchange): The hospital highlighted the success of free influenza vaccination for adults aged 60+, which significantly reduced infections in this group. Similarly, they stressed adult vaccination against shingles for those 50+, especially those with chronic conditions.
• Effect (The Impact): We refined our target demographic to explicitly include older adults and immunocompromised individuals, who are often excluded from live-virus vaccines but are ideal candidates for our CRISPRa-based approach.
• Adjust (The Refinement): We are now designing age-stratified in vitro models, and even though the results won’t be included in the competition, we hope to conduct more research into how advanced cellular age impacts the gene response that we get from CRISPRa.

Cycle 3: Combating Multiple Pathogens – Beyond Single-Virus Focus

• Acknowledge (The New Problem): Our system was originally designed with a narrow focus on SARS-CoV-2 and influenza-like viruses due to their recent relevance.
• Research (The Exchange): The hospital report noted that patients may face co-circulation of influenza, MPN, RSV, and other pathogens—emphasizing the need for broad-spectrum preparedness.
• Effect (The Impact): We decided to move away from singular gene induction, and instead, try and combine different sets of genes in order to produce an antiviral effect at multiple stages of pathogenesis, upping the degree to which we modulate the immune system.
• Adjust (The Refinement): We are now incorporating multiplexed sgRNAs into our LNP-hydrogel system to simultaneously upregulate multiple host defense pathways, increasing the breadth of protection.

Conclusion

By integrating real-world vaccination strategies and clinical perspectives from Hangzhou First People’s Hospital, we have refined our CRISPRa platform to better align with public health needs—broadening its timing, target population, pathogen coverage, and durability. These adjustments enhance the practical relevance and potential impact of our system as a novel, gene-based alternative to seasonal vaccines, particularly for high-risk and elderly populations.