Abstract: Objective This study aims to investigate the specific regulatory effect of miR-4454 on ZIKV replication and reveal its potential molecular mechanisms, providing new ideas and targets for the treatment of ZIKV infection. Methods Cells were transfected with miR-4454 mimics or inhibitors and subsequently infected with ZIKV. The expression of ZIKV NS5 mRNA and NS1 protein was measured using qRT-PCR and Western blot analyses. Bioinformatics tools were employed to predict potential target genes of miR-4454, focusing on the validation of LRPAP1 (low-density lipoprotein receptor-related protein associated protein 1) as a downstream target. Following the transfection of LRPAP1 plasmid or si-LRPAP1 into cells, ZIKV infection was performed, and qRT-PCR was used to detect ZIKV NS5 mRNA and interferon-stimulated gene expression, while Western blotting assessed the levels of ZIKV NS1 protein, IFNAR1, and phosphorylated STAT1, to explore the effect of LRPAP1 expression changes on ZIKV replication and the Type Ⅰ interferon signaling pathway. Results Overexpression of miR-4454 in cells inhibited the replication of ZIKV NS5 mRNA and expression of NS1 protein. Bioinformatics prediction identified LRPAP1 as a potential target of miR-4454, which is related to viral infection and the Type Ⅰ interferon signaling pathway. Dual-luciferase reporter assays confirmed that LRPAP1 is a direct target of miR-4454. Overexpression of LRPAP1 promoted ZIKV replication and suppressed the Type Ⅰ interferon signaling pathway, while knockdown of LRPAP1 inhibited ZIKV replication and activated this signaling pathway. Conclusion miR-4454 modulates ZIKV infection through the intrinsic immune pathway, suggesting its potential as a therapeutic target and providing new insights for the development of related treatment strategies.

Key words: Zika virus, Non-coding RNA, MiR-4454, Type Ⅰ interferon signaling pathway