The DOCK2 Knockout NCI-H1975 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population with targeted disruption of the DOCK2 gene in the NCI-H1975 lung adenocarcinoma cell line. This loss-of-function model is provided as a heterogeneous pool, which avoids clonal bias and is suitable for bulk functional assays such as migration and invasion studies. The polyclonal format preserves genetic diversity, offering a robust tool for investigating DOCK2-dependent processes without the need for single-cell cloning.
NCI-H1975 is a human lung adenocarcinoma cell line derived from a female nonsmoker, bearing EGFR L858R and T790M mutations. These mutations drive constitutive EGFR signaling and confer resistance to first-generation tyrosine kinase inhibitors, making the line a relevant model for EGFR-mutant non-small cell lung cancer. The cells retain key tumorigenic properties, including anchorage-dependent growth and metastatic potential, and are widely used in cancer biology research. This background provides a clinically relevant context for studying DOCK2 function in tumor cell motility.
DOCK2 encodes a Rac-specific guanine nucleotide exchange factor that operates in complex with ELMO1. It is activated downstream of chemokine receptors (e.g., CXCR4, CCR7) and the T cell receptor via PI3K and Src family kinases. Once stimulated, the DOCK2?CELMO1 complex catalyzes GTP loading on Rac1, which then engages effectors such as PAK, WAVE2, and the Arp2/3 complex to drive actin polymerization and lamellipodia formation. Interacting proteins like CrkL and Vav fine-tune this signaling cascade, positioning DOCK2 as a critical convergence point for cues that regulate directed cell migration.
In NCI-H1975 cells, DOCK2 knockout impairs Rac1-mediated actin cytoskeletal remodeling, resulting in diminished migration and invasion. This phenotype highlights DOCK2’s role in cancer metastasis and provides a direct model for studying NSCLC dissemination. Moreover, because DOCK2 is essential for immune cell trafficking, the knockout cells can be used in co-culture experiments to explore tumor-immune cell interactions. The model thus enables dissection of both cancer-autonomous and non-autonomous roles of DOCK2 in the tumor microenvironment.
This polyclonal knockout population is suitable for a range of assays to probe cell motility and signaling. Wound healing and transwell migration assays measure collective and single-cell migration, while Matrigel invasion assesses matrix degradation. Rac1 activation pull-downs and phospho-PAK western blotting reveal downstream signaling events. Immunofluorescence for F-actin visualizes cytoskeletal organization, and chemotaxis assays quantify response to chemoattractants like CXCL12. These applications make the model ideal for anti-metastatic drug screening and mechanistic studies. For technical inquiries, please contact Ascent Research.
