The HERC4 Knockout HEK293T Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T human embryonic kidney epithelial cell line. This polyclonal population is generated by CRISPR/Cas9-mediated gene disruption of the HERC4 locus, creating a loss-of-function model for studying HERC4-dependent biological processes. The polyclonal format ensures genetic heterogeneity while maintaining robust knockout across the population, making it suitable for bulk assays and high-throughput screening applications. This product is ideal for researchers investigating the ubiquitin-proteasome system, antiviral innate immunity, and ferroptosis regulation.
The parental HEK293T cell line originates from human embryonic kidney cells immortalized by adenovirus 5 DNA and constitutively expressing the SV40 large T antigen. This genetic background confers high transfection efficiency and robust protein expression capabilities, establishing HEK293T as a standard host for recombinant protein production, lentiviral packaging, and signaling pathway analysis. The cells retain epithelial morphology and key signaling networks of kidney-derived cells, providing a physiologically relevant context for studying ubiquitination dynamics and cellular stress responses. Their ease of culture and genetic manipulability make them a versatile platform for loss-of-function studies using CRISPR technology.
HERC4 encodes an E3 ubiquitin-protein ligase that catalyzes ubiquitin transfer to substrates, targeting them for proteasomal degradation. It negatively regulates antiviral innate immunity by ubiquitinating MAVS, an adaptor protein in the RIG-I pathway, leading to MAVS degradation and suppression of NF-??B signaling downstream of viral sensing. HERC4 also promotes ferroptosis by ubiquitinating GPX4, a key lipid peroxidase inhibitor, thereby sensitizing cells to iron-dependent oxidative death. Upstream, HERC4 expression is activated by type I interferons (IFN-??/??) via JAK-STAT signaling involving STAT1 and STAT2, and by IRF3 and IRF7 transcription factors during viral infection. Thus, HERC4 links ubiquitination to immune modulation and ferroptosis.
In the HEK293T context, HERC4 knockout allows precise examination of its role in ubiquitin-mediated regulation without interference from variable endogenous expression. The knockout model is especially useful for dissecting how HERC4-dependent MAVS degradation controls interferon induction and NF-??B-driven inflammatory responses. Additionally, HEK293T cells are responsive to ferroptotic triggers, enabling study of GPX4 turnover and lipid peroxidation pathways relevant to cancer and neurodegeneration. The polyclonal population avoids clonal artifacts, ensuring detectable and consistent phenotypes across a genetically diverse cell pool.
This product supports a range of assays including western blotting for ubiquitination and target protein levels, co-immunoprecipitation for interaction analysis, RT-qPCR for antiviral gene profiling, and flow cytometry for ferroptosis or viability assessment. Applications span viral pathogenesis, cancer biology, and neurodegenerative disease research focused on ubiquitin-proteasome system dysfunction. NF-??B and interferon reporter assays further enable functional interrogation of MAVS-dependent signaling. The HERC4 Knockout HEK293T Polyclonal Cells provide a robust platform for HERC4 research. For customized support, contact Ascent Research.
