The ABCF3 Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population derived from the human embryonic kidney HEK293T cell line, engineered for disruption of the ABCF3 gene. This product provides a heterogeneous cell pool with targeted gene disruption, enabling loss-of-function studies of ABCF3. The cells are offered as a ready-to-use knockout model for investigating immune signaling and translational control, leveraging the established utility of HEK293T cells in biomedical research.
HEK293T cells, a widely utilized host line, are human embryonic kidney cells transformed with adenovirus type 5 DNA and stably expressing the SV40 large T-antigen. This genetic modification enhances episomal replication and supports high levels of transient protein expression. The cells exhibit rapid proliferation, ease of transfection, and are a preferred platform for gene editing, CRISPR screening, and innate immune pathway analysis. Their robust nature and well-characterized biology make them suitable for generating polyclonal knockout pools.
ABCF3 belongs to the ATP-binding cassette transporter family and functions primarily as a negative regulator of antiviral innate immunity. Biochemically, ABCF3 interacts with the adaptor protein TANK and disrupts the STING?CTBK1?CIRF3 signaling axis. Under normal conditions, ABCF3 restrains type I interferon production by inhibiting TBK1-mediated phosphorylation of IRF3 and subsequent transcriptional activation of IFN-?? and interferon-stimulated genes. Upstream of ABCF3, innate immune stimuli such as cytosolic DNA or RNA activate STING, which in turn recruits TBK1, while downstream effectors IRF3 and NF-??B drive cytokine expression. By suppressing this cascade, ABCF3 limits excessive inflammatory responses; its knockout relieves this inhibition, resulting in potentiated STING-dependent signaling.
In the HEK293T background, ablation of ABCF3 creates a cell model with derepressed innate immune signaling pathways. This is particularly significant because HEK293T cells are known to have some intrinsic deficiencies in certain innate immune components, yet can be reconstituted for pathway studies. The ABCF3 knockout therefore allows researchers to study how removal of a key inhibitory checkpoint amplifies antiviral responses, providing a cleaner background for assaying STING/TBK1/IRF3 activation. This model is instrumental for dissecting negative feedback mechanisms, identifying pathway modulators, and exploring the interplay between translation control and immune defense.
Researchers can employ this knockout cell population in a variety of assays, including RT-qPCR quantification of IFN-?? and ISG transcripts, western blot analysis of phospho-TBK1 and phospho-IRF3 levels, luciferase reporter assays driven by the IFN-?? promoter, and co-immunoprecipitation studies of the ABCF3?CTANK interaction. Additional applications encompass RNA sequencing to profile global transcriptional changes, cytokine secretion measurements via ELISA, viral infection assays to evaluate replication kinetics, and flow cytometry to monitor immune activation markers. This polyclonal knockout model is suitable for fundamental studies in viral infection, cancer immunotherapy, and inflammatory disorder research, as well as for screening small molecules that target the STING pathway. For further technical inquiries, please contact Ascent Research.
