CASP4 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of the CASP4 gene. As a polyclonal pool, this reagent provides a functional loss-of-expression model without clonal selection, capturing a range of editing outcomes. The product abrogates CASP4 protein function for dissecting innate immune roles in lung adenocarcinoma.
The NCI-H1975 cell line is a widely used human non-small cell lung cancer model derived from the lung adenocarcinoma of a non-smoking female patient. These adherent epithelial cells harbor activating EGFR mutations L858R and T790M, conferring sensitivity and acquired resistance to first-generation EGFR tyrosine kinase inhibitors, respectively. NCI-H1975 cells are extensively employed in cancer biology and drug development for studying EGFR-targeted therapies and resistance mechanisms.
CASP4 is an inflammatory caspase that acts as an intracellular LPS receptor, binding the lipid A moiety to trigger autoproteolytic activation. Activated CASP4 initiates the non-canonical inflammasome pathway, cleaving GSDMD to generate N-terminal fragments that form plasma membrane pores and induce pyroptosis. This lytic cell death releases mature IL-1??, IL-18, and HMGB1. CASP4 expression is induced by type I interferons, NF-??B, and IRF1, and crosstalk with the NLRP3 inflammasome may amplify inflammation. Key signaling components include CASP4, LPS, GSDMD, IL-1??, IL-18, and NLRP3.
In the NCI-H1975 context, CASP4 knockout enables investigation of pyroptosis intersecting with EGFR oncogenic signaling. Lung adenocarcinoma cells often exhibit altered inflammatory responses influencing tumor progression and therapy. Removing CASP4-dependent pyroptosis allows examination of how immunogenic cell death impacts survival, cytokine profiles, and EGFR inhibitor sensitivity. This model also facilitates host?Cpathogen studies in a lung cancer-relevant background, given the role of bacterial infections in modulating tumor biology via innate immunity.
This polyclonal knockout pool supports diverse experimental approaches, including Western blotting and RT-qPCR for CASP4 and downstream targets GSDMD, IL-1??, and IL-18. Pyroptosis is assessed via LDH release, CellTiter-Glo viability, or IncuCyte imaging after intracellular LPS delivery. ELISA quantifies IL-1?? and IL-18 secretion. Combining LPS with EGFR inhibitors allows exploration of inflammatory contributions to TKI resistance. The model is suitable for rescue experiments to confirm pathway connections. For further information, please contact Ascent Research.
