This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human LoVo colorectal adenocarcinoma cell line, targeting the APOBEC3A gene. The polyclonal nature of this knockout model provides a heterogeneous pool of cells harboring disruptions in APOBEC3A, enabling robust functional studies without the bias of single-cell clonal selection. This loss-of-function tool is designed for researchers investigating the multifaceted roles of APOBEC3A in innate immunity and cancer mutagenesis.
The LoVo host cell line is a well-characterized epithelial model of metastatic colorectal adenocarcinoma, originally isolated from a supraclavicular lymph node metastasis. Notably, LoVo cells exhibit high-frequency microsatellite instability (MSI-H) and an intrinsic mutator phenotype, making them particularly relevant for studying DNA repair defects and mutation accumulation. Their tumorigenic properties and responsiveness to DNA-damaging agents further position LoVo as a versatile platform for oncology research.
APOBEC3A is a cytidine deaminase catalyzing C-to-U editing of ssDNA, acting as a restriction factor against retrotransposons (LINE-1, Alu) and viruses (HIV-1, hepatitis B). Its expression is regulated by interferon signaling, with upstream activators IFN-??, IFN-??, STAT1, IRF3, and NF-??B driving transcription upon pathogen detection. Once expressed, APOBEC3A targets viral cDNA and genomic retroelements, interacting with host factors such as APOBEC3G, HIV-1 Vif, UNG, SMUG1, and RPA. Downstream, UNG2 processes deaminated uracil, mediating mutagenic or antiviral outcomes. In cancer, aberrant APOBEC3A creates clustered kataegis mutations across the genome.
Ablating APOBEC3A function in the LoVo background creates a unique model for dissecting its contribution to the mutator phenotype inherent to MSI-H colorectal cancer cells. Loss of APOBEC3A-mediated deamination eliminates a source of C-to-T transitions and kataegic foci, thereby enabling researchers to isolate the impact of APOBEC3A on spontaneous and therapy-induced mutagenesis. Additionally, this knockout system facilitates investigation of APOBEC3A-dependant restriction of retrotransposon mobilization and viral infection in a cancer-relevant cellular environment, shedding light on the interplay between innate immunity and oncogenesis.
This polyclonal knockout cell population is suitable for a broad range of applications, including APOBEC mutational signature profiling via whole-genome sequencing, functional retrotransposon mobilization assays (e.g., LINE-1 retrotransposition reporter systems), and innate immune response studies following interferon stimulation or viral challenge. The cells can be employed in HIV-1 infection assays to assess viral restriction mechanisms or in drug sensitivity screens to evaluate DNA repair-targeted therapies. Researchers may also use techniques such as western blotting, RT-qPCR, immunofluorescence, and flow cytometry to validate pathway engagement and cellular phenotypes. For detailed technical specifications, additional validation data, or assistance with experimental design, please contact Ascent Research.
