APOBEC3F Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HEK293T cells, with targeted disruption of the APOBEC3F gene. This product contains a genetically heterogeneous pool of cells carrying gene edits, minimizing clonal bias and providing a versatile loss-of-function model. As a polyclonal population, it avoids the functional adaptation often associated with single-cell clones.
The HEK293T cell line is a human embryonic kidney epithelial line transformed by adenovirus 5 and constitutively expressing SV40 large T-antigen. Widely used for high-efficiency transfection and protein production, HEK293T cells serve as a standard platform for virology and functional genomics. The SV40 T-antigen also enables episomal replication of plasmids with SV40 origins, enhancing ectopic expression.
APOBEC3F is an interferon-inducible cytidine deaminase that restricts retroviruses and retrotransposons. Signaling through IFNAR1/IFNAR2 activates JAK1/TYK2 kinases, which phosphorylate STAT1 and STAT2; the activated heterodimer partners with IRF9 to bind ISRE elements and induce APOBEC3F transcription. APOBEC3F deaminates cytosine to uracil on single-stranded DNA, introducing lethal G-to-A hypermutation in HIV-1 cDNA during reverse transcription. Its activity is potently counteracted by HIV-1 Vif, which assembles a CBF-beta?Ccontaining ubiquitin ligase complex to target APOBEC3F for proteasomal degradation.
Although HEK293T cells lack certain T-cell factors, they provide a clean and tractable system for dissecting APOBEC3F-mediated restriction. The knockout background eliminates endogenous APOBEC3F activity, enabling unambiguous analysis of Vif function and APOBEC3F?CDNA editing events. This polyclonal population avoids clonal artifacts and is well-suited for studying interferon-regulated antiviral pathways and cancer-associated mutagenesis driven by APOBEC enzymes.
Applications include HIV-1 infectivity assays to quantify restriction, Western blotting and RT-qPCR for APOBEC3F expression analysis, and 3D-PCR for measuring mutation frequencies. Co-immunoprecipitation and immunofluorescence microscopy allow detection of interactions with Vif, CBF-beta, and RNA, while flow cytometry facilitates population-level studies. The model also supports drug screening targeting the APOBEC3F?CVif interface and investigations into APOBEC-imposed mutational signatures. For further details, contact Ascent Research.
