Cell culture-based influenza vaccines exhibit comparable safety and immunogenicity to
traditional egg-based vaccines. However, improving viral yield remains a key challenge in
optimizing cell culture-based production systems. Madin–Darby canine kidney (MDCK)
cells, the predominant cell line for influenza vaccine production, inherently activate interferon (IFN)-mediated antiviral defenses that restrict viral replication. To overcome
this limitation, we employed CRISPR/Cas9 gene-editing technology to generate an IFN
alpha/beta receptor subunit 1 (IFNAR1)-knockout (KO) adherent MDCK cell line. Viral titer analysis demonstrated significant enhancements in the yield of multiple vaccine
strains (H1N1, H3N2,... More
Cell culture-based influenza vaccines exhibit comparable safety and immunogenicity to
traditional egg-based vaccines. However, improving viral yield remains a key challenge in
optimizing cell culture-based production systems. Madin–Darby canine kidney (MDCK)
cells, the predominant cell line for influenza vaccine production, inherently activate interferon (IFN)-mediated antiviral defenses that restrict viral replication. To overcome
this limitation, we employed CRISPR/Cas9 gene-editing technology to generate an IFN
alpha/beta receptor subunit 1 (IFNAR1)-knockout (KO) adherent MDCK cell line. Viral titer analysis demonstrated significant enhancements in the yield of multiple vaccine
strains (H1N1, H3N2, and type B) in IFNAR1-KO cells compared to wild-type (WT) cells.
Transcriptomic profiling revealed marked downregulation of key interferon-stimulated
genes (ISGs)—including OAS, MX2, and ISG15—within the IFNAR1-KO cells, indicating a
persistent suppression of antiviral responses that established a more permissive microenvironment for influenza virus replication. Collectively, the engineered IFNAR1-KO cell
line provides a valuable tool for influenza virus research and a promising strategy for
optimizing large-scale MDCK cell cultures to enhance vaccine production efficiency.
