The APOBEC3A Knockout SK-OV-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the SK-OV-3 human ovarian adenocarcinoma cell line. This product provides a heterogeneous pool of APOBEC3A-disrupted cells, enabling functional studies free from clonal artifacts. The CRISPR/Cas9-mediated gene disruption abolishes APOBEC3A cytidine deaminase activity, creating a loss-of-function model suited for investigating the gene??s role in cancer biology.
SK-OV-3 is a well-characterized cell line established from the ascites of a patient with metastatic ovarian adenocarcinoma. It harbors oncogenic mutations in TP53 and KRAS, and exhibits adherent, epithelial growth. This genetic background supports investigations into tumor progression, genomic instability, and therapeutic resistance, making it an ideal host for studying APOBEC3A-mediated mutagenesis in ovarian cancer.
APOBEC3A is a cytidine deaminase that targets single-stranded DNA, inducing C-to-U edits primarily at TCA/TCT motifs. Its expression is induced by interferon-??/?? signaling via JAK1/TYK2-mediated phosphorylation of STAT1 and IRF1, with additional modulation by inflammatory cytokines TNF-?? and IL-1??. APOBEC3A interacts with replication protein A (RPA) to access ssDNA substrates, and its activity provokes DNA damage responses involving ATM, ATR, CHK1, and CHK2. Uracil DNA glycosylase UNG2 processes the resulting uracil bases, potentiating mutagenic outcomes. Consequently, APOBEC3A activity fuels APOBEC signature mutations and drives genomic instability.
In the SK-OV-3 background, with its mutant p53 and KRAS, APOBEC3A knockout enables dissection of how deaminase-driven mutation accumulation intersects with defective DNA repair and oncogenic signaling. This model is valuable for examining APOBEC3A??s contribution to the mutational landscape of ovarian tumors, particularly in the context of platinum-based chemotherapy resistance. Without APOBEC3A, researchers can determine baseline mutation rates and distinguish APOBEC-dependent from APOBEC-independent mutagenic processes.
Typical applications include quantifying APOBEC-specific C-to-T mutations via whole-exome or whole-genome sequencing, performing 3D-PCR to detect low-frequency editing, and using reporter assays to measure residual deaminase activity. The polyclonal nature supports population-scale studies of mutation dynamics and drug response heterogeneity. Additional uses involve probing APOBEC3A??s role in antiviral innate immunity and investigating its link to chronic inflammation-associated mutagenesis. Standard validation methods include western blotting for APOBEC3A and downstream DNA damage markers, comet assays, and RNA-seq. For further information, please contact Ascent Research.
