KCTD15 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the KCTD15 gene in the human NCI-H1975 lung adenocarcinoma cell line. This polyclonal format consists of a heterogeneous pool of cells harboring diverse editing outcomes, collectively leading to disrupted KCTD15 function without the need for single-cell cloning. The product provides a robust loss-of-function model for investigating the molecular roles of KCTD15 in a physiologically relevant non-small cell lung cancer (NSCLC) background. It is suitable for a wide range of functional genomics, signaling pathway dissection, and drug screening applications.
NCI-H1975 is a well-characterized human lung adenocarcinoma epithelial cell line derived from a patient with NSCLC. It carries activating EGFR L858R and T790M mutations, which drive constitutive oncogenic signaling and confer resistance to first-generation EGFR tyrosine kinase inhibitors. This genetic background makes the cell line a valuable model for studying EGFR-mutant lung adenocarcinoma, particularly in the context of acquired drug resistance and tumor progression. The cells exhibit stable epithelial morphology and reliable growth characteristics, enabling reproducible in vitro assays including proliferation, apoptosis, and pharmacological sensitivity testing.
KCTD15 encodes a substrate adaptor for the Cullin3-RING E3 ubiquitin ligase (CRL3) complex, where it interacts directly with CUL3 and RBX1 to mediate ubiquitination and proteasomal degradation of target proteins. Key substrates include histone deacetylases HDAC1 and HDAC2, linking KCTD15 to epigenetic regulation and transcriptional control. Functioning downstream of GLI transcription factors, KCTD15 modulates Hedgehog signaling by promoting turnover of these HDACs, thereby influencing gene expression programs that govern cell cycle progression, apoptosis, and neural crest development. Additionally, KCTD15 activity is regulated by TGF-?? signaling components, placing it at the intersection of multiple pathways critical for cellular homeostasis and oncogenesis.
In the EGFR-mutant NCI-H1975 context, KCTD15 knockout offers a powerful tool for dissecting the interplay between ubiquitin-mediated proteolysis, Hedgehog signaling, and lung cancer cell survival. The constitutive EGFR activity in this line provides an opportunity to evaluate how loss of KCTD15 alters downstream signaling networks, epigenetic landscapes, and drug response. Given emerging evidence of Hedgehog pathway involvement in NSCLC maintenance and therapy resistance, this polyclonal knockout pool enables investigation of tumor-intrinsic vulnerabilities and potential synthetic lethal interactions. The heterogeneous nature of the polyclonal population may also permit exploration of clonal dynamics within a cancer cell population.
This product supports diverse experimental applications. Ubiquitination and proteasome inhibition assays can directly monitor KCTD15 substrate turnover, while co-immunoprecipitation studies examine CRL3 complex formation. Western blotting and RT-qPCR quantify effects on HDAC1/2 protein levels and Hedgehog target gene expression. Cell proliferation, apoptosis, and flow cytometry-based cell cycle analyses assess functional consequences of knockout on growth and survival. Drug sensitivity profiling against EGFR inhibitors or Hedgehog antagonists may reveal novel therapeutic targets, and RNA-seq provides global transcriptional insights. For additional information, please contact Ascent Research.
