The JAK2 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the CAL-27 human tongue squamous cell carcinoma line, designed to disrupt the JAK2 gene. This pooled loss-of-function model maintains population heterogeneity, avoiding clonal biases, and is ideal for studying JAK2-dependent signaling in head and neck cancer. It enables robust in vitro assays to dissect oncogenic mechanisms and evaluate targeted therapeutics.
CAL-27 is an epithelial cell line isolated from a 56-year-old male with tongue squamous cell carcinoma, widely employed as a model for head and neck squamous cell carcinoma (HNSCC). The line exhibits aggressive growth and invasive traits, and JAK2 signaling is often dysregulated in HNSCC, associated with tumor progression and therapy resistance. Thus, JAK2 disruption in CAL-27 provides a relevant system to examine its contributions to malignant phenotypes.
JAK2 is a non-receptor tyrosine kinase that binds to cytokine receptors such as gp130 and EPOR. Upon ligand stimulation, JAK2 becomes activated and phosphorylates STAT transcription factors, notably STAT1, STAT3, and STAT5, which then dimerize and translocate to the nucleus to regulate target genes. JAK2 also engages the PI3K-AKT pathway by phosphorylating AKT and the MAPK/ERK cascade by activating ERK1/2. These pathways promote the expression of pro-survival and proliferative proteins including BCL-2, cyclin D1, and c-MYC. The signaling strength is modulated by negative regulators like SOCS1, SOCS3, SHP-1, and SHP-2, which interact with and inhibit JAK2, ensuring tight control of cellular responses to cytokines.
In the CAL-27 background, JAK2 knockout is expected to dampen oncogenic signaling. Since JAK2 drives transcription of anti-apoptotic BCL-2 and cell cycle promoters cyclin D1 and c-MYC, its loss should impair proliferation and survival. Additionally, JAK2-mediated phosphorylation of STAT3 and AKT enhances motility and invasion, so knockout cells likely show reduced migration. This model thus enables assessment of JAK2 dependency in HNSCC, informing the therapeutic potential of JAK inhibition in this disease.
Researchers can use this polyclonal knockout population for western blot analysis of JAK2/STAT pathway components, RT-qPCR for downstream target genes, and phospho-STAT flow cytometry. Functional assays such as MTT proliferation, transwell migration/invasion, and Annexin V apoptosis assays quantify the phenotypic consequences of JAK2 loss. The model is suitable for drug screening with JAK inhibitors like ruxolitinib and for investigating synergy with other targeted agents. It also supports co-culture studies to explore the role of JAK2 in tumor-immune interactions. For further information, please contact Ascent Research.
