The CD2AP Knockout NCI-H1975 Polyclonal Cells provide a heterogeneous CRISPR/Cas9-edited population of NCI-H1975 human lung adenocarcinoma epithelial cells carrying targeted disruptions in the CD2AP gene. This polyclonal knockout model enables the study of CD2AP-dependent functions without the clonal selection artifacts often present in single-cell-derived lines, offering a robust tool for investigating adaptor protein biology in a non-small cell lung cancer (NSCLC) background.
The parental NCI-H1975 cell line is a widely utilized model for NSCLC, derived from a patient with lung adenocarcinoma and harboring activating mutations in EGFR (L858R/T790M) and PIK3CA (G118D). These genetic lesions drive oncogenic signaling through the PI3K/AKT and MAPK pathways, making the cell line particularly valuable for examining the interplay between receptor tyrosine kinase signaling and cytoskeletal regulation. The polyclonal knockout pools generated from this background retain the genomic complexity of the original tumor-derived line while ablating CD2AP expression.
CD2AP encodes an essential intracellular adaptor protein that bridges membrane receptors, such as CD2 and nephrin, to the actin cytoskeleton and endocytic machinery. It functions downstream of multiple upstream regulators, including EGFR and Src family kinases activated by PI3K-mediated signals, and directly interacts with cortactin (CTTN), the p85 regulatory subunit of PI3K, and actin. Through these interactions, CD2AP orchestrates actin polymerization dynamics and endosomal trafficking by recruiting factors like Rab4 and Rab5. Consequently, CD2AP loss disrupts cortactin-mediated actin remodeling and impairs receptor-mediated endocytosis, potentially altering downstream effectors such as Akt and mTOR.
In lung adenocarcinoma, CD2AP has been implicated in tumor progression, with some evidence suggesting a tumor-suppressive function through its regulation of cell adhesion and migration. Knockout in the NCI-H1975 line, which already exhibits intrinsic invasive properties due to EGFR and PIK3CA mutations, allows researchers to dissect the contribution of CD2AP to NSCLC phenotypes. The polyclonal cell population is especially suited for assessing heterogeneous responses to CD2AP ablation, such as changes in collective cell migration, invasion capacity, and sensitivity to targeted therapies. Furthermore, because CD2AP is a critical component of the glomerular filtration barrier and its mutation causes focal segmental glomerulosclerosis (FSGS), this model can also be adapted to explore molecular mechanisms of podocyte biology.
Typical experimental applications include assessing cell migration and invasion via wound healing and transwell assays, analyzing signaling pathway alterations by phospho-Akt western blotting, and examining protein interactions through co-immunoprecipitation of cortactin or p85. Proliferation and drug sensitivity studies can be conducted to evaluate the impact of CD2AP loss on NSCLC therapeutic responses, while immunofluorescence microscopy permits visualization of actin cytoskeleton reorganization. RT-qPCR and western blotting are recommended for confirming CD2AP knockout efficiency in the polyclonal population. For more information on this product and custom gene editing services, please contact Ascent Research.
