The APOBEC3G Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney epithelial cell line. This loss-of-function model disrupts the APOBEC3G gene, a cytidine deaminase that restricts HIV-1 replication. The polyclonal format provides a robust, genetically heterogeneous pool of edited cells, enabling reproducible studies of APOBEC3G-mediated antiviral activity and its regulation.
The HEK293T host cells stably express the SV40 large T antigen, facilitating episomal replication of plasmids with SV40 origins and conferring high transfection efficiency. This cell line is widely employed for protein expression, virus production, and functional genomics due to its rapid growth and ease of manipulation. Its well-characterized biology and genetic tractability make it an ideal platform for generating knockout models to investigate human innate immune factors.
APOBEC3G inhibits HIV-1 replication by deaminating viral cDNA, inducing G-to-A hypermutations that impair reverse transcription. Its expression is transcriptionally activated by type I interferons through the JAK-STAT pathway, involving IFNAR1/2, JAK1, TYK2, STAT1, STAT2, and IRF9 forming the ISGF3 complex. To evade this restriction, HIV-1 Vif recruits a Cullin5?CElongin B/C?CRbx2 E3 ubiquitin ligase, with the cofactor CBF-??, targeting APOBEC3G for proteasomal degradation. APOBEC3G also interacts with related deaminases APOBEC3F and APOBEC3H.
HEK293T cells do not express APOBEC3G endogenously, so this knockout eliminates any inducible background, providing a defined system for reconstitution experiments. The model is ideal for dissecting Vif-mediated degradation by reintroducing wild-type or mutant APOBEC3G constructs. The cells’ high transfection efficiency supports robust HIV-1 vector production and single-cycle infectivity assays, enabling high-throughput screening for compounds that disrupt the Vif?CAPOBEC3G interaction. The polyclonal format maintains genetic heterogeneity while ensuring consistent loss of function.
These knockout cells enable a range of assays, including HIV-1 single-cycle infectivity comparisons to quantify APOBEC3G restriction, 3D-PCR for hypermutation detection, and co-immunoprecipitation of Vif?CAPOBEC3G complexes for degradation pathway analysis. RT-qPCR of interferon-stimulated genes assesses innate immune activation, while immunofluorescence and viral replication kinetics monitor antiviral responses. Beyond virology, the model is applicable to APOBEC3G-mediated cancer mutagenesis studies and drug screening for Vif inhibitors. For further details or to inquire about this product, please contact Ascent Research.
