The CBLL1 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1299 human lung adenocarcinoma cell line, with targeted disruption of the CBLL1 gene. This polyclonal format encompasses a heterogeneous mix of loss-of-function mutations, providing a robust experimental system that reflects tumor genetic diversity without the constraints of clonal selection. The product is supplied as a ready-to-use knockout pool ideal for investigating CBLL1 function in cancer cell biology and epithelial-mesenchymal transition (EMT) research.
NCI-H1299 is a widely utilized human non-small cell lung cancer (NSCLC) cell line originally isolated from a lymph node metastasis of a 43-year-old male with lung adenocarcinoma. These adherent epithelial cells are p53-deficient and exhibit a highly metastatic phenotype, making them a clinically relevant in vitro platform for studying tumor invasion and dissemination. The p53-null background further emphasizes pathways that drive malignancy and provides a permissive environment for examining oncogenic signaling and therapeutic responses.
CBLL1 functions as an E3 ubiquitin ligase that specifically targets E-cadherin for ubiquitin-mediated proteasomal degradation following Src kinase phosphorylation. This degradation is activated by upstream signals such as EGF, HGF, and Wnt, and involves direct interaction with c-Src, phosphotyrosine proteins, and E2 ubiquitin-conjugating enzymes. Loss of E-cadherin leads to adherens junction disassembly, release of ??-catenin and p120-catenin, and subsequent actin cytoskeleton remodeling. These events culminate in the transcriptional induction of Snail and Slug, master regulators of EMT that drive cellular migration and invasion.
In the NCI-H1299 cell context, genetic ablation of CBLL1 is predicted to stabilize E-cadherin at the plasma membrane, restore cell?Ccell adhesion, and attenuate the intrinsically high migratory and invasive capacity associated with p53 loss. This model offers a powerful tool for dissecting the reliance of metastatic NSCLC on CBLL1-driven EMT. The polyclonal knockout population also enables assessment of functional heterogeneity that may emerge when ubiquitin ligase activity is disrupted in a metastatic tumor background.
A diverse array of experimental applications is supported, including transwell migration and invasion assays to quantify metastatic potential, immunofluorescence microscopy to visualize E-cadherin and ??-catenin localization, and ubiquitination assays to probe CBLL1 enzymatic activity. Co-immunoprecipitation experiments can dissect interactions with Src and E2 enzymes, while RT-qPCR and western blotting enable profiling of EMT markers such as Snail and Slug. Cell viability and proliferation studies can assess the impact of CBLL1 loss on tumor growth, and the model is amenable to high-throughput screening for EMT or Src inhibitors. For further assistance, contact Ascent Research.
