The CCDC71L Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of the CCDC71L gene. This heterogeneous knockout pool retains genetic diversity while eliminating the need for clonal isolation, enabling rapid phenotypic screening without clonal bias and providing a robust platform for loss-of-function studies.
NCI-H1299 is a human lung adenocarcinoma cell line established from a lymph node metastasis of a patient with non-small cell lung cancer. Adherent and epithelial in morphology, these cells maintain invasive and migratory characteristics of the metastatic phenotype, making them a widely used tumorigenic model for studying lung cancer progression and therapeutic responses.
CCDC71L encodes a protein featuring predicted coiled-coil domains, motifs commonly associated with protein scaffolding and protein-protein interactions. Its molecular function remains uncharacterized, but it is hypothesized to influence cytoskeletal organization and cell adhesion through putative interactions with actin filaments, microtubules, and focal adhesion complexes. Disruption of CCDC71L likely alters cytoskeletal dynamics, cell morphology, and migratory capacity. Given its coiled-coil architecture, CCDC71L may form homo- or heterodimers with other coiled-coil proteins, potentially acting as a scaffolding hub that integrates cytoskeletal and adhesion signals. Potential signaling connections may place CCDC71L downstream of integrin-mediated adhesion or upstream of actin polymerization, though specific binding partners and regulators have not been defined.
Knockout of CCDC71L in the metastatic NCI-H1299 background provides a relevant model for probing the role of scaffold proteins in lung cancer cell migration and invasion. The polyclonal format minimizes clonal adaptation artifacts, permitting assessment of average phenotypic effects across diverse editing events. This model is therefore valuable for functional genomics efforts aimed at identifying genes that modulate the aggressiveness of lung adenocarcinoma.
Typical applications include wound healing and transwell migration/invasion assays to evaluate motility changes, immunofluorescence staining of actin and focal adhesions for cytoskeletal analysis, and proliferation assays to measure growth alterations. Knockout validation is performed via western blot and RT-qPCR to confirm protein and mRNA reduction, respectively. Global transcriptomic profiling by RNA-seq can uncover downstream pathway perturbations. For further details or technical assistance, please contact Ascent Research.
