The APOBEC3A Knockout HT29 Polyclonal Cells provide a loss-of-function model for studying the cytidine deaminase APOBEC3A in a colorectal adenocarcinoma background. Generated by CRISPR/Cas9-mediated gene disruption, this polyclonal cell population harbors heterogeneous editing events at the APOBEC3A locus, eliminating functional protein expression. The product is intended for research into innate immunity, APOBEC-mediated mutagenesis, and colorectal cancer.
The HT29 cell line is a well-established human colorectal adenocarcinoma model derived from a primary tumor, widely used to study intestinal epithelial biology and colorectal cancer. HT29 cells form polarized monolayers and tight junctions, enabling investigations of cell signaling and oncogenic transformation. This knockout model allows examination of APOBEC3A function within an epithelial tumor context.
APOBEC3A is a cytidine deaminase that catalyzes cytosine-to-uracil deamination on single-stranded DNA, generating C-to-T mutations that drive genomic instability. It is transcriptionally induced by interferon-alpha/beta through the IFNAR receptor, leading to JAK1/TYK2-mediated phosphorylation and activation of STAT1, which together with IRF1 promotes APOBEC3A expression. At single-stranded DNA, APOBEC3A interacts with RPA and PCNA to introduce lesions that are recognized by uracil DNA glycosylases UNG and SMUG1. Unrepaired damage can activate a TP53-dependent DNA damage response, linking innate immune signaling to mutagenesis and apoptosis.
In HT29 cells, APOBEC3A knockout reduces APOBEC-driven C-to-T mutational signatures and may alter responsiveness to DNA-damaging therapies. This model facilitates dissection of APOBEC3A??s contributions to colorectal cancer mutagenesis and the crosstalk between interferon signaling and DNA repair pathways, offering insights into tumor evolution and immune-driven genomic changes.
Applications include next-generation sequencing to map APOBEC-specific mutations, RT-qPCR and western blotting for interferon-stimulated gene analysis, ??H2AX-based DNA damage assays, and functional studies of cell viability, migration, and invasion. For additional information or technical support, please contact Ascent Research.
