The DTX4 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the DTX4 gene in the HAP1 cell model. This loss-of-function model enables systematic interrogation of DTX4-dependent regulatory mechanisms without generating a clonal isolate, providing a heterogeneous gene-edited pool suitable for functional genomics and population-level phenotypic analyses. The product is a ready-to-use reagent for researchers aiming to investigate the role of DTX4 in innate immune signaling, ubiquitin-mediated proteolysis, and related pathways.
The HAP1 host cell line is a near-haploid derivative of the KBM-7 chronic myeloid leukemia cell line, broadly employed for CRISPR-based knockout and functional genomics studies due to its stable karyotype and ease of genetic manipulation. HAP1 cells retain key components of innate immune signaling cascades, making them a well-characterized platform for dissecting antiviral responses and interferon regulation. Their near-haploid nature minimizes confounding effects of multiple alleles, facilitating consistent and interpretable knockout phenotypes in polyclonal populations.
DTX4 encodes an E3 ubiquitin ligase that acts as a negative regulator of type I interferon signaling by catalyzing K48-linked ubiquitination of TBK1, targeting it for proteasomal degradation. This modification limits TBK1-mediated phosphorylation of IRF3, thereby attenuating IFN-?? and interferon-stimulated gene (ISG) expression. DTX4 function is activated by viral infection, TLR3/TLR4 signaling, and IFN-?? itself, placing it within a negative feedback circuit. Additional interactors include TRAF3 and NOTCH1, linking DTX4 to both antiviral and Notch signaling networks. By controlling TBK1 turnover, DTX4 modulates the amplitude and duration of antiviral innate immune responses.
In the HAP1 chronic myeloid leukemia background, disruption of DTX4 is expected to enhance TBK1 stability and prolong IRF3 activation, potentially boosting IFN-?? production and antiviral gene expression. This model provides a unique context to study how interferon regulatory dynamics intersect with leukemogenesis and immune evasion. The polyclonal nature of the knockout population allows researchers to assess functional heterogeneity and dominant effects without clonal selection bias, which is particularly valuable when exploring immune-modulatory pathways that may influence cell fitness and proliferation.
This knockout product supports a wide range of applications, including innate antiviral immunity studies, negative regulation of interferon signaling, Notch pathway dissection, and E3 ubiquitin ligase characterization. Typical downstream assays include Western blotting for TBK1 and IRF3 to assess protein stability and phosphorylation states, RT-qPCR for IFN-?? and ISGs to quantify transcriptional responses, ubiquitination and proteasomal degradation assays to monitor TBK1 turnover, co-immunoprecipitation for interaction studies, antiviral response luciferase reporter assays, and phospho-signaling flow cytometry. The DTX4 Knockout HAP1 Polyclonal Cells thus serve as a versatile tool for dissecting ubiquitin-dependent immune regulation and for drug discovery campaigns focused on modulating interferon responses. For further details, please contact Ascent Research.
