The APOBEC3A Knockout NCI-H1299 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout pool featuring targeted disruption of the APOBEC3A gene in the NCI-H1299 human lung adenocarcinoma cell line. This polyclonal population provides a versatile loss-of-function model, circumventing the limitations of single-cell clones while enabling robust analysis of APOBEC3A-mediated biological processes.
The NCI-H1299 parental cell line originates from a metastatic lymph node of a lung adenocarcinoma patient, bearing a KRAS mutation and p53 deficiency. These genetic lesions promote genomic instability and aberrant signaling, establishing NCI-H1299 as a widely adopted model for studying oncogenic pathways, tumor progression, and DNA damage responses in non-small cell lung cancer.
APOBEC3A is a cytidine deaminase that edits single-stranded DNA, catalyzing C-to-U conversions at TCW motifs and generating somatic mutations in cancer genomes. Its expression is activated by interferon-alpha, interferon-gamma, DNA damage stimuli, and viral nucleic acids through NF-??B signaling. APOBEC3A interacts with UNG, APOBEC3B, and viral proteins such as HIV-1 Vif and HPV E6/E7. Its enzymatic activity induces mutations in TP53 and PIK3CA, triggers DNA double-strand breaks, and activates the DNA damage response. In innate immunity, APOBEC3A feeds into the cGAS-STING pathway, driving IRF3-dependent IFN?? transcription.
In NCI-H1299 cells, APOBEC3A knockout eliminates cytidine deaminase activity, reducing C-to-U mutations and APOBEC signature mutagenesis. This ablation diminishes genomic instability, potentially attenuating DNA damage signaling and altering innate immune pathways. The model isolates APOBEC3A??s role in promoting tumor heterogeneity, clonal evolution, and drug resistance, while providing a clean background to study mutagenic enzyme interactions with DNA repair networks.
Representative applications include dissecting APOBEC3A-mediated mutagenesis in lung adenocarcinoma, profiling DNA damage responses via ??H2AX foci, and performing drug sensitivity screens. The cells can be analyzed by Western blotting, RT-qPCR, and whole-genome sequencing for mutation signature analysis, as well as interferon response assays measuring STAT1 phosphorylation. For additional details, please contact Ascent Research.
