The KIF2A Knockout NCI-H1975 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population targeting the KIF2A gene in the human NCI-H1975 lung adenocarcinoma cell line. This pooled knockout model is generated by CRISPR/Cas9-mediated disruption of the KIF2A locus, resulting in a heterogeneous population of cells with loss-of-function mutations. Unlike monoclonal cell lines, this polyclonal format preserves the genetic diversity of the edited pool, which can better reflect subtle phenotypic variability and reduce clonal bias in functional studies. The product provides a robust tool for investigating KIF2A-dependent processes in a non-small cell lung cancer (NSCLC) background without requiring isolation of a single clone.
NCI-H1975 is a widely characterized human lung adenocarcinoma cell line derived from the pleural effusion of a 55-year-old Caucasian female nonsmoker with NSCLC. This cell line harbors notable genetic alterations, including an EGFR T790M/L858R double mutation, rendering it a clinically relevant model for studying EGFR-targeted therapies and drug resistance mechanisms. NCI-H1975 cells exhibit epithelial morphology and are commonly employed in cancer biology research to dissect signaling pathways governing tumor growth, metastasis, and therapeutic response. Integration of the KIF2A knockout into this genetic background offers a unique system to explore interactions between microtubule dynamics, mitotic regulation, and oncogenic signaling in a defined lung adenocarcinoma context.
KIF2A encodes a kinesin-13 family microtubule depolymerase that catalyzes ATP-dependent depolymerization of microtubule plus ends, a function critical for mitotic spindle assembly, chromosome congression, and cytokinesis. Its activity is tightly controlled by upstream kinases including Aurora A (AURKA), Aurora B (AURKB), Polo-like kinase 1 (PLK1), and cyclin-dependent kinase 1 (CDK1), which phosphorylate KIF2A to modulate its localization and depolymerase activity. KIF2A interacts with core microtubule regulators such as tubulin, EB1, CLASP1, and KIF2C/MCAK, and forms complexes with HSET and KBP. Beyond mitosis, KIF2A regulates ciliogenesis through interactions with CEP164 and influences downstream Hippo pathway effectors YAP1 and TAZ, thereby linking microtubule dynamics to transcriptional programs. The tumor suppressor TP53 has also been implicated as an upstream modulator, adding another layer of regulatory control in cancer cells.
Disruption of KIF2A in NCI-H1975 cells is expected to impair mitotic progression, leading to spindle assembly defects, chromosome missegregation, and prolonged mitosis, which can activate the spindle assembly checkpoint and potentially induce apoptosis. Given the critical role of microtubule dynamics in EGFR-driven lung cancers??particularly in mediating resistance to microtubule-targeting agents such as paclitaxel??this knockout model enables dissection of how KIF2A-dependent microtubule remodeling contributes to drug response. Additionally, loss of KIF2A may disrupt ciliogenesis-dependent YAP/TAZ signaling, which is known to regulate epithelial-mesenchymal transition (EMT) and stemness in NSCLC. Thus, the polyclonal KIF2A knockout population serves as a versatile tool to investigate how aberrant microtubule regulation intersects with oncogenic signaling in lung adenocarcinoma.
This product is ideally suited for applications in cancer cell biology, including detailed analysis of mitotic progression, chromosome instability, and ciliogenesis using methodologies such as immunofluorescence (??-tubulin, centromere staining), live-cell imaging of mitotic dynamics, and ciliogenesis assays. The knockout cells can be employed in drug sensitivity screens to evaluate responses to microtubule-targeting agents (e.g., paclitaxel) or Aurora kinase inhibitors, as well as in migration and invasion assays coupled with qRT-PCR or Western blotting to assess EMT and YAP/TAZ activity. Flow cytometry enables high-content analysis of cell cycle perturbations, while metaphase spread assays provide direct readouts of chromosome segregation fidelity. For technical inquiries or to discuss scaling or customized applications, please contact Ascent Research.
