The APOBEC3A Knockout PaTu 8988t Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human pancreatic ductal adenocarcinoma cell line PaTu 8988t. This loss-of-function model targets the cytidine deaminase APOBEC3A, a critical enzyme in innate immunity and cancer mutagenesis. The polyclonal format maintains genetic diversity while ablating APOBEC3A function, avoiding clonal selection biases and enabling robust study of APOBEC3A-dependent processes.
The PaTu 8988t cell line is derived from a liver metastasis of human pancreatic ductal adenocarcinoma, serving as an epithelial cancer model with high metastatic potential. It retains hallmark features of advanced pancreatic cancer, including aggressive growth and genomic instability. Its metastatic origin makes it particularly suitable for studying tumor dissemination and colonization mechanisms. Paired with APOBEC3A knockout, this line provides a platform to examine the consequences of APOBEC3A mutagenesis in a disease-relevant context.
APOBEC3A is a single-stranded DNA cytidine deaminase that deaminates cytidine to uridine, functioning in innate antiviral immunity while also inflicting somatic mutations across cancer genomes. Its expression is potently induced by interferons (IFN-??, -??, -??) signaling through the IFNAR1/2?CJAK1/TYK2?CSTAT1/2 axis, with cooperative activation by IRF9, NF-??B, IRF3, and IRF7. Once expressed, APOBEC3A edits genomic DNA, mitochondrial DNA, viral cDNA, and certain mRNAs, generating uracil lesions processed by uracil DNA glycosylase (UNG) that trigger DNA damage responses mediated by ATR, Chk1, and H2AX phosphorylation. The enzyme interacts with replication protein A (RPA), p53, and TCF4, connecting its mutagenic activity to replication stress and genome integrity surveillance.
In PaTu 8988t pancreatic cancer cells, APOBEC3A promotes a mutational signature of C-to-T and C-to-G substitutions in TpCpN motifs. Knocking out APOBEC3A in this polyclonal population is expected to eliminate this mutational process, reducing heterogeneity that drives tumor evolution, drug resistance, and metastasis. Moreover, since APOBEC3A-induced DNA lesions can activate interferon responses and inflammatory signaling, this model allows dissection of how endogenous mutagenesis impacts innate immune sensing and the tumor microenvironment. Thus, it serves as a tool to study the interplay between genome instability and immune evasion in metastatic pancreatic cancer.
This knockout model supports diverse applications, including tumor evolution studies via whole-genome sequencing and APOBEC mutation signature analysis, drug resistance profiling with clonogenic survival and dose-response assays, and innate immune sensing investigations through RT-qPCR and western blotting. DNA damage response may be assessed by ??-H2AX immunofluorescence. The polyclonal population is conducive to clonal dynamics and heterogeneity studies under selection. For further information or custom uses, please contact Ascent Research.
