Background: The emergence of the JN1 variant amid the COVID-19 pandemic has raised concerns regarding vaccination uptake. Understanding the factors influencing public perception and response to COVID-19 vaccination concerning this variant is critical. Methods: A mixed-methods approach was employed to investigate factors impacting COVID-19 vaccination acceptance. Surveys and interviews were conducted among diverse demographic groups to assess attitudes, beliefs, and concerns surrounding JN1-specific vaccination. Results: Findings revealed demographic variations in vaccine acceptance, with higher education levels and older age groups exhibiting greater acceptance. Trust in healthcare authorities, perceived vaccine efficacy, and concerns about side effects significantly influenced vaccination attitudes. Accessibility issues and the impact of misinformation were identified as barriers to uptake. Conclusion: Tailored strategies addressing concerns about vaccine safety and efficacy, countering misinformation, and improving accessibility are crucial to enhance vaccination uptake amidst the evolving JN1 variant.