biostudies-literatureCheng YTaizhou Science and Technology Support ProjectTaizhou Sixth Phase “311 High-level Talent Training Special Program”Taizhou Science and Technology Support Project (Social development)complementary funding project of Jiangsu Agri-Animal Husbandry Vocational CollegeScience and Technology Innovation Team Project of Jiangsu Agri-Animal Husbandry Vocational CollegeQing Lan Project of Jiangsu Province230https://www.ebi.ac.uk/biostudies/studies/S-EPMC12709809biostudies-literatureUnknown56(1)Duck Tembusu virus (DTMUV) is a single-stranded positive-sense RNA virus that poses a serious threat to the poultry industry, and its infection mechanism remains incompletely understood. This study aimed to investigate the role of the host protein DDX17 (DEAD-box RNA helicase 17) in DTMUV infection. Utilizing an in vitro infection model combined with CRISPR/Cas9 gene knockout, siRNA interference, and overexpression techniques, we found that DTMUV infection significantly upregulated both mRNA and protein expression levels of DDX17 in host cells. Furthermore, DDX17 facilitated viral replication through its ATP-binding and hydrolysis domain. Co-immunoprecipitation and mass spectrometry analysis confirmed an interaction between DDX17 and the DTMUV C protein. Additionally, phylogenetic analysisVeterinary researchDTMUV upregulates DDX17 expression to facilitate viral replication.PMC12709809NSFPT202518TS202405NSF2023TC02Wang MWu ZXie WMa CZhou QWang AZhang JJia WSun FZhou FCheng YZhu SfalseDTMUV upregulates DDX17 expression to facilitate viral replication.Duck Tembusu virus (DTMUV) is a single-stranded positive-sense RNA virus that poses a serious threat to the poultry industry, and its infection mechanism remains incompletely understood. This study aimed to investigate the role of the host protein DDX17 (DEAD-box RNA helicase 17) in DTMUV infection. Utilizing an in vitro infection model combined with CRISPR/Cas9 gene knockout, siRNA interference, and overexpression techniques, we found that DTMUV infection significantly upregulated both mRNA and protein expression levels of DDX17 in host cells. Furthermore, DDX17 facilitated viral replication through its ATP-binding and hydrolysis domain. Co-immunoprecipitation and mass spectrometry analysis confirmed an interaction between DDX17 and the DTMUV C protein. Additionally, phylogenetic analysis2025-01-01T00:00:00Z2025 Dec2026-05-26T20:01:52.438Z2026-05-25T03:12:05.238ZS-EPMC127098094140285710.1186/s13567-025-01639-0