〔Abstract〕 Objective To screen the RNA binding proteins interacting with NBV nonstructural protein σNS in host cells and to analyze their bioinformatics functions. Methods In this study, the eukaryotic expression vector pEF-HA-MB-S3 of NBV σ NS was constructed and transfected into HEK293T cells after verification. After RNase A treatment, the obtained protein lysate was enriched by immunoprecipitation to enrich σNS binding proteins, identified and analyzed by LC-MS/MS mass spectrometry, and the properties and specific functions of proteins were discovered with the help of related Bioinformatics tools. Results In this study, 32 candidate RNA binding proteins interacting with NBV σNS proteins were successfully screened, and the results of bioanalysis showed that these proteins were mainly located in cytoplasm and nucleus, and were mainly involved in biological processes such as cell metabolism, biological regulation, virus translation and transcription. Conclusion This study preliminarily analyzes the function of RNA binding proteins interacting with NBV σNS, which lays a foundation for further study on the mechanism of σNS protein in NBV life cycle.