The IVNS1ABP Knockout NCI-H1975 Polyclonal Cells are a genetically modified human cell population generated by CRISPR/Cas9-mediated disruption of the IVNS1ABP gene in the NCI-H1975 lung adenocarcinoma epithelial cell line. This product is supplied as a polyclonal knockout cell pool, providing a heterogeneous population of edited cells for loss-of-function studies. The targeted gene disruption eliminates functional IVNS1ABP protein expression, enabling researchers to investigate the regulatory roles of this actin-binding adaptor in innate immune signaling pathways. The polyclonal format maintains cellular diversity and is well-suited for bulk functional assays, including western blotting, RT-qPCR, and viral infection experiments, without requiring single-cell clonal isolation.
The host cell line, NCI-H1975, is derived from the lung adenocarcinoma of a 62-year-old female and harbors oncogenic mutations in EGFR (L858R) and TP53 (R273H). These cells serve as a widely validated model for non-small cell lung cancer (NSCLC), particularly for studying EGFR-driven tumorigenesis and therapeutic resistance. The epithelial morphology and adherent growth characteristics of NCI-H1975 cells facilitate standard cell culture techniques and high-content imaging assays. The presence of endogenous EGFR and TP53 mutations provides a clinically relevant genetic background for exploring interactions between oncogenic signaling and innate immune regulation.
IVNS1ABP (influenza virus NS1A binding protein) functions as a critical negative regulator of antiviral innate immunity by bridging viral sensors and downstream adaptors. Mechanistically, IVNS1ABP binds to the adaptor protein TRIF and disrupts TLR3- and RIG-I-mediated signaling cascades, thereby suppressing the activation of TBK1 and the transcription factors IRF3 and NF-??B. This inhibition culminates in reduced transcriptional induction of type I interferons, including interferon-beta, and pro-inflammatory cytokines. IVNS1ABP is upregulated by interferon-alpha, interferon-beta, and STAT1/STAT2-containing transcriptional complexes, establishing a negative feedback loop. Additionally, IVNS1ABP interacts with the influenza A virus NS1 protein and cytoskeletal regulators such as actin and filamin A, linking innate immune signaling to cytoskeletal dynamics.
Knockout of IVNS1ABP in the NCI-H1975 background is expected to potentiate TLR3 and RIG-I signaling, leading to enhanced IRF3 and NF-??B activation, elevated interferon-beta production, and heightened expression of interferon-stimulated genes. This hyperactive innate immune phenotype renders the model particularly valuable for dissecting host?Cpathogen interactions, especially during influenza A virus infection, where viral NS1 normally exploits IVNS1ABP to dampen antiviral responses. Furthermore, the EGFR-mutant lung adenocarcinoma environment provides a unique platform to examine the crosstalk between oncogenic MAPK/STAT pathways and innate immunity, with potential implications for understanding immune evasion in NSCLC and evaluating immune-modulating therapeutic strategies.
Researchers can employ these polyclonal knockout cells in a broad range of experimental contexts. Representative applications include western blot analysis of phosphorylated and total IRF3, NF-??B, and STAT1; RT-qPCR quantification of IFN-?? and downstream interferon-stimulated gene transcripts; and viral infection assays with influenza A to assess replication kinetics and host defense. Additional assays such as immunofluorescence staining for NF-??B nuclear translocation, phospho-signaling analysis via phospho-TBK1 and phospho-IRF3 detection, and dual luciferase reporter assays for NF-??B and IFN-?? promoter activity provide versatile tools for mechanistic studies. This model supports investigations in innate immune signaling, viral pathogenesis, cancer immunology, and drug screening for immune modulators. For further technical inquiries, please contact Ascent Research.
