CCDC91 Knockout NCI-H1299 Polyclonal Cells represent a ready-to-use CRISPR/Cas9-edited polyclonal knockout cell population designed for functional investigation of the CCDC91 gene in a human non-small cell lung carcinoma background. These polyclonal cells are produced by introducing a CRISPR/Cas9-mediated disruption of the target gene in the NCI-H1299 host cell line, generating a heterogeneous pool of loss-of-function alleles without single-cell cloning. This format preserves genetic diversity while providing a robust model for studying CCDC91-dependent processes in a population context, ideal for assays that require consistent knockout phenotypes across a cell bulk rather than clonal expansion.
The NCI-H1299 cell line is a widely employed model of invasive lung adenocarcinoma, originally established from a lymph node metastasis. It harbors a homozygous TP53 mutation and wild-type EGFR, making it particularly relevant for studies of p53-deficient metastatic progression and resistance to EGFR-targeted therapies. NCI-H1299 cells exhibit pronounced migratory and invasive capabilities in vitro, and they are frequently utilized to dissect the molecular mechanisms of cancer cell motility, epithelial-to-mesenchymal transition (EMT), and extracellular matrix remodeling. This genetic and phenotypic context renders NCI-H1299 an optimal host for interrogating CCDC91 function in a disease-relevant setting.
CCDC91 encodes a coiled-coil domain protein that localizes to the Golgi apparatus and is critical for maintaining Golgi stack architecture and facilitating reorientation toward the leading edge during directional cell migration. Mechanistically, CCDC91 interacts with the Golgi stacking protein GRASP55 (GORASP2) and cis-Golgi matrix protein GM130 (GOLGA2) to organize Golgi structure. It also associates with syntaxin 5, a SNARE protein involved in vesicle trafficking. Upstream, the Rho GTPase CDC42, a master regulator of cell polarity, activates protein kinases that control Golgi dynamics, thereby positioning CCDC91 within a CDC42?CGRASP55?CGM130 signaling axis. Disruption of CCDC91 impairs the repositioning of the Golgi ribbon, leading to altered directed secretion of extracellular matrix remodeling factors, including matrix metalloproteinases (MMPs), which in turn affects focal adhesion turnover and cell motility.
In the NCI-H1299 background, knockout of CCDC91 provides a powerful tool to dissect the interplay between Golgi organization and metastatic behavior. Given the host cell??s intrinsic invasive capability and p53-deficient status, loss of CCDC91 is expected to compromise Golgi reorientation, reducing directional migration and invasion through matrigel or endothelial barriers. This model permits investigation of how Golgi-dependent secretion modulates the tumor microenvironment and facilitates cancer cell dissemination. Furthermore, it enables exploration of the molecular connections between oncogenic signaling, Golgi remodeling, and EMT-driven cell plasticity, potentially revealing new vulnerabilities in metastatic lung adenocarcinoma.
Researchers can employ this polyclonal knockout product in a wide range of experimental workflows. Common applications include high-resolution immunofluorescence analysis of Golgi morphology using antibodies against GM130 and GRASP55 to assess structural integrity and polarization. Functional migration and invasion can be measured via wound healing assays and transwell chambers, while co-immunoprecipitation experiments validate the CCDC91?CGRASP55 interaction and identify additional binding partners. Quantitative Western blotting for MMP-2 and MMP-9 secretion provides readouts of downstream Golgi-dependent secretion. Additionally, these cells are suitable for high-throughput screening campaigns designed to discover pharmacological modulators of Golgi dynamics, and for Rho GTPase activity assays (e.g., CDC42 activation) to link upstream signaling to CCDC91 function. For further information or to discuss custom genome engineering projects, please contact Ascent Research.
