The APOBEC3C Knockout HEK293T Polyclonal Cells from Ascent Research represent a CRISPR/Cas9-edited polyclonal population of HEK293T cells in which the APOBEC3C gene has been disrupted to create a versatile loss-of-function model. The polyclonal format provides a heterogeneous pool of edited cells that collectively ensure robust target-gene disruption while minimizing clonal artifacts. This product is designed for researchers investigating APOBEC3C-mediated innate antiviral immunity, DNA deamination, and associated mutagenic processes in a human cellular context.
HEK293T is a widely employed human embryonic kidney epithelial cell line engineered to stably express the SV40 large T antigen. This expression enables episomal replication of plasmids containing the SV40 origin, markedly enhancing transient transfection efficiency and protein yield. The cell line is an established workhorse for viral vector production, CRISPR screening, and heterologous protein expression, offering high transfectability, rapid growth, and compatibility with diverse functional assays.
APOBEC3C functions as a cytidine deaminase that transcriptionally responds to type I interferon signaling through IRF3, IRF7, and the ISGF3 complex comprising STAT1, STAT2, and IRF9. Upon activation, the enzyme deaminates cytosine to uracil in single-stranded DNA intermediates of retroviruses and retrotransposons, causing lethal G-to-A hypermutation. APOBEC3C is counteracted by the HIV-1 Vif protein, which recruits the enzyme to the Cullin5-ElonginB-ElonginC E3 ubiquitin ligase complex for proteasomal degradation. Key substrates include HIV-1, HBV, and HPV viral genomes, as well as LINE-1 and Alu retroelements; off-target activity on host genomic DNA contributes to APOBEC signature mutations observed in various cancers.
In HEK293T cells, which lack robust endogenous expression of many antiviral factors, the APOBEC3C knockout provides a clean background for dissecting its specific contributions to retroviral restriction and DNA mutagenesis. This system enables interrogation of APOBEC3C in isolation, avoiding functional overlap with other APOBEC3 family members, and is particularly suited for mechanistic studies of the Vif-Cullin5 ubiquitin ligase axis and mapping of deamination targets in a human kidney epithelial environment.
Typical experimental applications include HIV-1 infectivity assays, LINE-1 retrotransposition reporter systems, and cytidine deaminase activity measurements, often combined with RT-qPCR or Western blotting for knockout validation. The product is also valuable for cancer mutagenesis studies, where APOBEC3C-specific mutation signatures can be resolved through next-generation sequencing. For more information, please contact Ascent Research.
