This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human non-small cell lung carcinoma cell line, featuring targeted disruption of the HERC1 gene. The polyclonal format comprises a heterogeneous pool of edited cells, providing a robust loss-of-function model without clonal selection artifacts. This gene-edited population enables exploration of HERC1-dependent mechanisms in a therapeutically relevant epithelial cancer background.
The host NCI-H1975 cell line is a lung adenocarcinoma model established from a 70-year-old female. It harbors an activating PIK3CA mutation (H1047R) while remaining wild-type for EGFR and KRAS, reflecting a molecular subtype responsive to PI3K pathway inhibitors. These epithelial cells are widely employed in cancer biology and preclinical drug response studies, making them an ideal platform for investigating tumor suppressors and oncogenic signaling networks.
HERC1 encodes a large E3 ubiquitin ligase with guanine nucleotide exchange factor (GEF) activity for small GTPases. Mechanistically, HERC1 ubiquitinates ARF6 and clathrin, thereby controlling membrane trafficking events including endocytosis and autophagy. Its activity is regulated by upstream factors such as ARF6 and CDK1, and it interacts with NEURL4, HERC2, RAB35, and 14-3-3 proteins. Through these interactions, HERC1 influences downstream processes such as cell cycle progression via cyclin B/CDK1, mTOR signaling through ULK1 and ATG5, and protein homeostasis by targeting substrates to the ubiquitin-proteasome system. Potential downstream effectors include p53 and SMAD2/3, linking HERC1 to genome stability and TGF-?? responses.
In the NCI-H1975 context, HERC1 knockout likely disrupts endosomal trafficking and autophagic flux, processes often dysregulated in lung adenocarcinoma. The PIK3CA-mutant background may synergize with loss of HERC1 to alter mTOR pathway activity, cell cycle distribution, and sensitivity to PI3K/mTOR inhibitors. This model thus provides a relevant system to dissect how ubiquitin-mediated membrane dynamics intersect with oncogenic signaling, potentially revealing vulnerabilities in epithelial cancers.
This polyclonal knockout cell population is suitable for a wide range of functional assays. Researchers can assess HERC1-dependent changes in protein ubiquitination, autophagy markers (e.g., LC3 lipidation), and cell cycle profiles via flow cytometry. Standard applications include proliferation, migration, and clonogenic assays, as well as drug sensitivity screens with agents such as PI3K or mTOR inhibitors. Co-immunoprecipitation and immunofluorescence experiments can probe altered subcellular trafficking of ARF6 or clathrin. These cells also support RT-qPCR and Western blotting for pathway validation. For further technical details, please contact Ascent Research.
