The IFIH1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from HeLa cervical adenocarcinoma cells, featuring targeted disruption of the IFIH1 gene. This polyclonal format yields a heterogeneous pool of knockout genotypes, avoiding clonal bias and providing a robust model for loss-of-function studies in pooled screening and population-level assays. The cells are supplied cryopreserved and can be expanded for experiments requiring consistent genetic background across replicates.
HeLa cells are a human epithelial line isolated from an HPV18-positive cervical adenocarcinoma, characterized by stable adherent growth and p53 inactivation via HPV E6 oncoprotein. They are permissive to diverse viral pathogens and widely used in virology and cancer research, offering extensive molecular annotations and high transfection efficiency. This background facilitates investigation of innate immune signaling without the confounding effects of primary cell variability.
IFIH1 encodes MDA5, a RIG-I-like receptor that detects long cytosolic double-stranded RNA and signals through the mitochondrial adaptor MAVS. Activated MAVS recruits TRAF3 and triggers TBK1/IKK?? kinases, which phosphorylate IRF3 and IRF7, leading to transcriptional induction of type I interferons (IFN-??/??) and downstream ISGs such as ISG15, OAS1, and PKR. The pathway is modulated by cofactors including ZCCHC3, TRIM25, and PACT, and dysregulation is linked to autoinflammatory disorders like Aicardi-Gouti??res syndrome.
In the HeLa context, IFIH1 disruption abrogates the primary cytoplasmic dsRNA sensing pathway, rendering cells deficient in MDA5-driven interferon and cytokine induction. This allows precise elucidation of RIG-I versus MDA5 substrate specificity, quantification of viral replication differences, and investigation of how HPV-associated p53 inactivation intersects with antiviral innate immunity. Consequently, the model aids in defining pathway bifurcations and feedback regulation under defined genetic perturbation.
Research applications include viral infection assays with EMCV or SARS-CoV-2, Poly(I:C) stimulation followed by IFN-?? and cytokine ELISA, RT-qPCR profiling of ISGs, and RNA-seq transcriptomics to map interferon-driven gene networks. The polyclonal pool is also suited for co-immunoprecipitation to assess disrupted MDA5?CMAVS interactions and for high-throughput screens to identify compounds that restore or bypass MDA5 signaling. For technical consultation, please contact Ascent Research.
