AIM2 Knockout HaCaT Cell Line

The AIM2 Knockout HaCaT Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the widely used HaCaT immortalized human keratinocyte cell line. This product features targeted disruption of the AIM2 gene, which encodes absent in melanoma 2, a cytosolic DNA sensor that initiates inflammasome assembly. CRISPR/Cas9-mediated gene disruption eliminates AIM2 protein expression, providing a loss-of-function model for studying innate immune pathways. The cell line is supplied as a validated knockout model, ready for use in advanced biomedical research on inflammatory skin diseases and DNA sensing mechanisms.

HaCaT cells are a spontaneously immortalized human keratinocyte cell line originally derived from adult skin tissue. These cells retain essential features of normal epidermal keratinocytes, including the capacity to differentiate and form stratified epithelial layers, while remaining non-tumorigenic. HaCaT cells are extensively employed in dermatological research as a model for skin barrier function, immune responses, and keratinocyte biology. Their well-characterized signaling networks make them an ideal host for gene knockout studies, particularly those investigating cutaneous innate immunity and inflammatory signaling pathways.

AIM2 functions as a critical pattern recognition receptor that detects double-stranded DNA in the cytoplasm. Upon ligand binding, AIM2 recruits the adaptor protein ASC/PYCARD, which oligomerizes to form ASC specks that serve as platforms for caspase-1 activation. Active caspase-1 then cleaves pro-IL-1?? and pro-IL-18 into their mature, secreted forms, and also processes gasdermin D to trigger pyroptosis. AIM2 expression is upregulated by type I interferons and interferon regulatory factors, integrating DNA sensing with broader innate immune activation. Additionally, AIM2 interacts with other sensor proteins such as IFI16. This signaling cascade culminates in the release of potent inflammatory cytokines and can induce immunogenic cell death.

In HaCaT keratinocytes, the AIM2 inflammasome mediates responses to cytosolic DNA derived from various sources, including microbial pathogens and cellular damage. Disruption of AIM2 in these cells abrogates inflammasome assembly upon DNA stimulation, leading to impaired ASC speck formation, reduced caspase-1 cleavage, and diminished secretion of IL-1?? and IL-18. This knockout model enables precise dissection of AIM2-dependent signaling from other innate pathways in epidermal cells. As keratinocytes are frontline sentinels in the skin, this cell line is particularly relevant for studying the roles of DNA sensing in psoriasis, atopic dermatitis, skin infections, and autoinflammatory disorders. It also provides a platform to investigate pyroptosis regulation and its impact on skin barrier integrity.

The AIM2 Knockout HaCaT Cell Line is a versatile tool for research on inflammasome biology and inflammatory skin diseases. Typical applications include functional studies of the AIM2-ASC-caspase-1 axis following transfection of poly(dA:dT) or other DNA ligands. It facilitates screening of anti-inflammatory compounds, with assays such as ELISA for IL-1?? and IL-18, western blotting for caspase-1 cleavage, and immunofluorescence for ASC speck formation. Researchers can also employ this model to examine gasdermin D-mediated pyroptosis via LDH release and to explore crosstalk with interferon pathways. This knockout cell line contributes to drug discovery and mechanistic investigations in academic and industrial settings. For further information, please contact Ascent Research.