随着逆遗传技术的发展和对包装信号的理解提升，流感A病毒（IAV）用于病毒治疗和生物技术的应用大大加快。虽然使用复制能力型IAV尤其具有潜力，因其高感染力能够有效传播至器官深部，但同时也存在着其无法以细胞特异性方式进行控制而引发传染病的风险。因此，本文开发了一个简单而有效的复制能力型IAV基于细胞靶向系统。实验证明，IAV的核糖核蛋白复合物（RNP）的活性可以通过内源蛋白和与RNA依赖性RNA聚合酶亚单位融合的纳米抗体之间的相互作用来调节。为了验证该方法的可行性，实验证明了包含与p53相对的纳米抗体Nb139融合聚合酶的RNP在表达内源p53的HEK293T细胞中是不活跃的，但在p53缺陷的Saos-2细胞中是活跃的。最后，成功制备了只能在p53缺陷的肿瘤细胞中进行繁殖的复制能力型IAV，以及利用细胞特异性方式递送外源基因的IAV载体。该方法具有灵活性，因为纳米抗体可以很容易地改变以靶向不同的细胞类型，为病毒治疗和生物技术提供了一个有价值的平台。版权所有 © 2023. Elsevier B.V.出版

Applications of influenza A viruses (IAV) for virotherapy and biotechnology have accelerated substantially with the development of reverse genetic technology and advances in the understanding of packaging signals. While the use of a replication-competent IAV is particularly promising, owing to its efficient transmission to organ depths with high infectivity, there is also a risk that its multiplication cannot be controlled in a cell-type-specific manner, causing an infectious disease. Therefore, here a simple and effective replication-competent IAV-based cell-targeting system has been developed. It was demonstrated that the activity of the ribonucleoprotein complex (RNP) of IAV could be regulated by the interaction between the endogenous protein and a nanobody fused to the subunit of RNA-dependent RNA polymerase (RdRp). To validate the feasibility of the method, it was demonstrated that RNP containing RdRp fused with Nb139, a nanobody against p53, is inactive in HEK293T cells expressing endogenous p53, but active in p53-defective Saos-2 cells. Finally, a replication-competent IAV was successfully generated that multiplies only in p53-defective tumor cells and an IAV vector was developed that can deliver a foreign gene in cell type-specific manner. The method is flexible because the nanobody can be easily altered to target a different cell type, offering a valuable platform for virotherapy and biotechnology.Copyright © 2023. Published by Elsevier B.V.