The APOBEC3A Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human renal adenocarcinoma line 786-O. This product provides a genetically disrupted APOBEC3A locus, creating a loss-of-function model for investigating the multifunctional roles of APOBEC3A. The polyclonal nature of the knockout pool enables population-level studies without clonal selection artifacts, and the knockout is validated at the population level.
The 786-O cell line is a widely used epithelial model of clear cell renal cell carcinoma (ccRCC). Established from a primary renal adenocarcinoma, it harbors a loss-of-function mutation in the von Hippel-Lindau (VHL) tumor suppressor gene, leading to stabilization of hypoxia-inducible factors (HIFs) and constitutive activation of hypoxic signaling. This VHL-deficient background mimics a key driver event in ccRCC, driving aberrant angiogenesis, glycolysis, and tumor progression. 786-O cells are adherent, maintain a stable karyotype, and are amenable to genetic manipulation, making them a standard platform for renal cancer research.
APOBEC3A functions as a single-stranded DNA cytidine deaminase, converting cytosines to uracils and generating U:G mismatches. These are processed by UNG and APE1, causing DNA breaks that activate a DDR involving gamma-H2AX, ATM, ATR, and p53. APOBEC3A transcription is induced by interferon signaling: interferon-alpha/beta and interferon-gamma activate JAK1/TYK2, which phosphorylate STAT1/STAT2; these together with IRF9 form ISGF3 that binds APOBEC3A gene regulatory elements. NF-??B also upregulates APOBEC3A. APOBEC3A interacts with TRIB3 and is targeted by viral factors like HIV-1 Vif and HPV E6, underscoring its innate immune function.
In the context of 786-O ccRCC cells, APOBEC3A knockout is particularly valuable for dissecting the contribution of APOBEC-mediated mutagenesis to renal cancer evolution. APOBEC3A-induced mutation signatures are prevalent in many cancers, including ccRCC. By eliminating endogenous APOBEC3A activity, this model allows researchers to distinguish between APOBEC-dependent and independent mutational processes in a VHL-null background. Moreover, because APOBEC3A links innate immune signaling to genomic instability, its knockout provides a clean system to study interferon-driven DNA damage responses without confounding deaminase activity. This model may also help clarify how APOBEC3A-mediated mutagenesis influences drug sensitivity and resistance mechanisms in renal cancer.
Typical applications include whole-genome sequencing for mutational signature analysis, DNA damage assays (gamma-H2AX immunofluorescence, comet assays), viral restriction experiments, and drug response profiling. Cells are validated by western blotting for APOBEC3A and RT-qPCR. For further information or to discuss custom applications, please contact Ascent Research.
