Introduction: Persistent infection with human papillomavirus 16 (HPV16) is one of the main etiologic causes of oropharyngeal squamous cell carcinoma (OPSCC). OPSCC is a rising problem around the world increasing in Asia, Europe, and North America. Our goal was to demonstrate whether CRISPR/Cas9 knock out of HPV16 E7 gene and adding a STING agonist treatment would result in effective tumour clearance and reactivation of the innate immune response. Methods: We evaluated the efficacy of collective treatment with a CRISPR/Cas9 inducible doxycycline system to edit HPV16 E7 and the effect of STING agonists in vitro and in vivo models. Results: In vitro models successfully knocked out the HPV16 E7 gene from OPSCC cells and combined with CT DNA as STING agonist, increased IFNβ. In vivo experiments showed that CRISPR of E7 in combination with STING agonist 2'3'-cGAMP produces a significant tumour regression and elevation of the IFNβ response. Conclusion: Joint therapy of CRISPR/Cas9 inducible system with STING agonists made possible the targeted inactivation of HPV16 E7, the significant regression of OPSCC tumours and the mitigation of the innate immune system response restricted by E7. This inducible knocking out system has the potential to be used as a powerful aid in immunotherapy treatments for OPSCC after adjusting for more specificity and less inflammatory responses.