Hantavirus (HV) infection, which underlies hantavirus hemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome, remains to be a severe clinical challenge. Here, we synthesized small interfering RNAs (siRNAs) that target the encoding sequences of HV strain 76-118, and validated their inhibitory role in virus replication in HV-infected monkey kidney Vero E6 cells. A chimeric protein, 3G1-Cκ-tP, consisting of a single-chain antibody fragment (3G1) against the HV surface envelop glycoprotein, the constant region of human immunoglobulin κ chain (Cκ), and truncated protamine (amino acids 8–29, tP), was further generated. The fusion protein showed high affinity to HV antigen on the infected cell membrane, and internalized through clathrin-mediated endocytosis; it bound to siRNAs via the basic nucleic acid-rich protamine fragment, leading to their specific delivery into HV-infected cells and efficient inhibition of virus replication. An encephalitis mouse model was established via intracranial HV administration. Intraperitoneal injection of siRNAs complexed with 3G1-Cκ-tP achieved specific distribution of siRNAs in HV-infected brain cells, significantly reduced HV antigen levels, and effective protection from HV infection-derived animal death. These results provide a compelling rationale for novel therapeutic protocols designed for HV infection and related disorders.

汉坦病毒（HV）感染是伴有肾综合征和汉坦病毒肺综合征的汉坦病毒出血热的基础，仍然是严峻的临床挑战。在这里，我们合成了靶向HV株76-118编码序列的小干扰RNA（siRNA），并验证了它们在HV感染的猴肾Vero E6细胞中的病毒复制中的抑制作用。嵌合蛋白3G1-Cκ-tP，由抗HV表面包膜糖蛋白的单链抗体片段（3G1），人免疫球蛋白κ链恒定区（Cκ）和截短的鱼精蛋白组成（氨基酸8-29， tP），进一步生成。融合蛋白对被感染的细胞膜上的HV抗原具有高亲和力，并通过网格蛋白介导的内吞作用而被内在化。它通过富含核酸的基本鱼精蛋白片段与siRNA结合，导致其特异性递送至HV感染的细胞并有效抑制病毒复制。通过颅内高压给药建立了脑炎小鼠模型。腹膜内注射与3G1-Cκ-tP复合的siRNA可在HV感染的脑细胞中实现siRNA的特异性分布，显着降低HV抗原水平，并有效防止HV感染引起的动物死亡。这些结果为针对HV感染和相关疾病而设计的新型治疗方案提供了令人信服的理由。显着降低了HV抗原水平，并有效防止了HV感染引起的动物死亡。这些结果为针对HV感染和相关疾病而设计的新型治疗方案提供了令人信服的理由。显着降低了HV抗原水平，并有效防止了HV感染引起的动物死亡。这些结果为针对HV感染和相关疾病而设计的新型治疗方案提供了令人信服的理由。