The HIP1R Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-mediated gene disruption model targeting HIP1R in a heterogeneous population of NCI-H1975 lung adenocarcinoma cells. This polyclonal knockout cell pool provides a physiologically relevant loss-of-function system for studying HIP1R-dependent processes without clonal selection artifacts. The use of CRISPR/Cas9 technology ensures targeted disruption of the HIP1R locus, and the polyclonal format preserves the genetic diversity of the parental line, enabling robust functional assays.
The NCI-H1975 cell line was derived from the pleural effusion of a non-smoking female with lung adenocarcinoma and carries EGFR L858R/T790M mutations along with a PIK3CA mutation, while retaining wild-type p53. This well-characterized line is a standard model for EGFR-mutant non-small cell lung cancer (NSCLC) and exhibits resistance to EGFR tyrosine kinase inhibitors, including osimertinib. Its epithelial origin and defined genetic background make it particularly suitable for studying endocytic trafficking, drug resistance mechanisms, and tumor cell migration in a clinically relevant context.
HIP1R functions as an adaptor linking clathrin-mediated endocytosis to the actin cytoskeleton. It is activated by EGFR signaling and serum growth factors, and interacts with clathrin, actin, huntingtin, cortactin, and the AP2 adaptor complex. HIP1R promotes clathrin-coated pit formation and actin filament bundling, facilitating EGFR internalization and degradation. Its downstream effects also influence cell migration through actin regulation. HIP1R knockout disrupts EGFR endocytosis, leading to altered signaling and potentially reduced invasion. This coordination of endocytic traffic and cytoskeletal remodeling is critical for receptor trafficking.
The HIP1R knockout in NCI-H1975 cells is particularly valuable given the EGFR-mutant background. EGFR L858R/T790M mutations drive constitutive signaling, which may be modulated by endocytic trafficking. Disrupting HIP1R allows dissection of how clathrin-mediated endocytosis influences EGFR signaling output, receptor half-life, and downstream pathways involved in proliferation and survival. The polyclonal nature of the knockout population provides a heterogeneous model that reflects the genetic and phenotypic variability observed in tumors, enabling studies on how HIP1R loss affects cell migration, invasion, and response to EGFR inhibitors like osimertinib. This model thus offers insights into resistance mechanisms and potential vulnerabilities in NSCLC.
Researchers can utilize these HIP1R knockout polyclonal cells in diverse functional assays, including EGFR degradation kinetics, transferrin uptake measurement to assess clathrin-mediated endocytosis, wound healing and Transwell invasion assays, and immunofluorescence staining for clathrin, cortactin, and actin. Moreover, these cells enable western blot analysis of EGFR and downstream signaling effectors, drug sensitivity profiling with osimertinib, co-immunoprecipitation to probe HIP1R interactors, and live-cell imaging to track endocytic dynamics. Such applications support investigations into EGFR trafficking, endocytosis-dependent signaling, and mechanisms of drug resistance and metastasis in lung adenocarcinoma. For further details or ordering information, please contact Ascent Research.
