The usefulness of live attenuated virus vaccines has been limited by suboptimal immunogenicity, safety concerns or cumbersome manufacturing processes and techniques. Here we describe the generation of a live attenuated influenza A virus vaccine using proteolysis-targeting chimeric (PROTAC) technology to degrade viral proteins via the endogenous ubiquitin–proteasome system of host cells. We engineered the genome of influenza A viruses in stable cell lines engineered for virus production to introduce a conditionally removable proteasome-targeting domain, generating fully infective PROTAC viruses that were live attenuated by the host protein degradation machinery upon infection. In mouse and ferret models, PROTAC viruses were highly attenuated and able to elicit robust and broad humoral, mucosal and cellular immunity against homologous and heterologous virus challenges. PROTAC-mediated attenuation of viruses may be broadly applicable for generating live attenuated vaccines.

减毒活病毒疫苗的用途受到次优免疫原性、安全问题或繁琐的制造过程和技术的限制。在这里，我们描述了使用蛋白水解靶向嵌合 (PROTAC) 技术通过宿主细胞的内源性泛素-蛋白酶体系统降解病毒蛋白来生成减毒甲型流感病毒活疫苗。我们在为病毒生产而设计的稳定细胞系中设计了甲型流感病毒的基因组，以引入有条件可移除的蛋白酶体靶向结构域，从而产生完全具有感染性的 PROTAC 病毒，这些病毒在感染后被宿主蛋白质降解机制活减毒。在小鼠和雪貂模型中，PROTAC 病毒高度减毒并能够引发强健和广泛的体液，针对同源和异源病毒挑战的粘膜和细胞免疫。PROTAC 介导的病毒减毒可能广泛适用于产生减毒活疫苗。