The HDDC2 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the HDDC2 gene. This product provides a heterogeneous pool of NCI-H1975 cells carrying loss-of-function edits, avoiding clonal selection and enabling pooled functional studies. It is optimized for molecular, biochemical, and cellular analyses in innate immune signaling and cancer biology research.
The parental NCI-H1975 cell line is a human lung adenocarcinoma epithelial model derived from pleural effusion. It harbors activating EGFR L858R and T790M mutations, driving constitutive kinase activity and TKI resistance. This widely used line models EGFR-mutant non-small cell lung cancer, facilitating studies on oncogenic signaling, drug resistance, and tumor?Cimmune interactions.
HDDC2 encodes an HD domain-containing phosphohydrolase that dephosphorylates 5??-triphosphate viral RNA, converting it to a 5??-diphosphate form and preventing RIG-I binding. This activity negatively regulates RIG-I?CMAVS aggregation and downstream signaling through TBK1, IKK??, IRF3, and NF-??B, thereby restraining IFN-?? and ISG15 induction. HDDC2 directly interacts with 5??-triphosphate RNA and is targeted by MERS-CoV papain-like protease. Its expression is itself upregulated by type I interferon, STAT1, and IRF3. CRISPR/Cas9-mediated knockout of HDDC2 removes this regulatory block, leading to enhanced RIG-I activation, IRF3 phosphorylation, and amplified type I interferon production upon viral RNA stimulation.
In the NCI-H1975 background, HDDC2 loss provides a model to study the crosstalk between innate antiviral pathways and EGFR-driven lung adenocarcinoma. Elevated RIG-I signaling can alter interferon responses and potentially reshape the tumor immune microenvironment. This system enables investigation of how antiviral signaling modulates tumor cell growth, cytokine secretion, or oncolytic virus sensitivity, offering insights relevant to immune-oncology.
Typical applications include measurement of RIG-I pathway activity by Western blotting, RT-qPCR, IFN-?? luciferase reporter assays, and ELISA; interrogation of IRF3 translocation via immunofluorescence and phospho-IRF3 flow cytometry; and functional evaluation using viral replication assays and RNA-seq. The cells support functional genomics screens, antiviral innate immunity research, and immune-oncology target validation. For further information, please contact Ascent Research.
