The CCDC14 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of the CCDC14 gene in the human NCI-H1299 lung carcinoma line. This polyclonal knockout model, generated via CRISPR/Cas9-mediated gene disruption, enables loss-of-function studies while maintaining population heterogeneity. It serves as a valuable tool for investigating CCDC14??s roles in ciliogenesis, protein?Cprotein interactions, and cancer cell biology without clonal selection artifacts.
NCI-H1299 is a non-small cell lung carcinoma cell line derived from a lymph node metastasis, featuring a homozygous TP53 deletion and wild-type KRAS. This genetic background provides a clinically relevant model of lung adenocarcinoma, widely used to study metastatic progression, p53-independent tumor biology, and drug resistance. The TP53-null status allows focused analysis of target gene disruptions unconfounded by intact p53 signaling.
CCDC14 encodes a coiled-coil domain protein implicated in ciliogenesis and cell cycle control. Primary cilia are essential for Hedgehog signaling, where SHH binding to PTCH1 derepresses SMO, activating GLI transcription factors (GLI1?C3). Although the exact interactors of CCDC14 remain undefined, its role in cilia formation suggests knockout may impair cilium assembly, thereby perturbing Hedgehog pathway transduction and downstream GLI-mediated transcription, offering a model to study cilia-dependent signaling networks.
In NCI-H1299 cells, CCDC14 disruption may compromise primary cilia and Hedgehog function, which are often co-opted in lung cancer progression. This knockout model therefore permits dissection of ciliogenesis contributions to tumorigenic properties, including proliferation, migration, and drug response, in a TP53-null, KRAS wild-type background. It provides a unique system to examine how loss of ciliary integrity influences metastatic behavior and ciliopathy-related mechanisms without confounding KRAS mutations.
Typical applications include immunofluorescence for cilia markers (acetylated tubulin, ARL13B), western blotting for target validation, MTT proliferation assays, and transwell migration studies to assess metastatic potential. Transcriptome analysis via RNA-seq can reveal global expression changes, while drug sensitivity screening with cisplatin or paclitaxel elucidates effects on chemoresistance. These assays support functional genomics of CCDC14 in NSCLC and ciliogenesis research. For further information, please contact Ascent Research.
