Introduction

CNS infections caused by flaviviruses, such as Japanese encephalitis virus (JEV) and West Nile viruses (WNV), lead to substantial neuroinflammation and neuronal apoptosis. Recent advancements in high-throughput sequencing techniques have revealed the emergence of numerous long non-coding RNAs (lncRNAs) as key regulators of viral infections. However, the precise role of lncRNAs in governing neuroinflammation and neuronal cell death during flavivirus infection remains inadequately understood. In our previous work, we demonstrated that the lncRNA JINR1 interacts with RNA binding protein (RBM10) and NF-κB, activating NF-κB target genes to enhance flavivirus-induced neuronal cell death and viral replication.

Methods

Cell culture, virus infection, titration, qRT-PCR, cloning and transfections, ChIP, Western blotting, and the dual luciferase assay were used.

Results and Conclusion

We found that JINR1 reduces the levels of mature, primary, and precursor miR-216b-5p in human neuronal cell lines. Mechanistically, JINR1 recruits H3K27me3 marks, reduces H3K4me3, and diminishes RNA polymerase-II recruitment at the miR-216b-5p promoter, thereby transcriptionally inhibiting the expression of miR-216b-5p. Furthermore, through dual-luciferase reporter assays, we demonstrated that JINR1 acts as a molecular sponge for miR-216b-5p, upregulating GRP78 expression through a competing endogenous RNA (ceRNA) regulatory mechanism. We are the pioneers in elucidating the regulatory function of JINR1, which involves suppressing the transcription of miR-216b-5p and serving as a ceRNA. Interestingly, we found that miR-216b-5p negatively regulates virus replication in a human neuronal cell line due to the presence of multiple binding sites in the viral genome.