The L1CAM Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 colorectal adenocarcinoma line, designed for loss-of-function studies of the human L1CAM gene. This product format consists of a heterogeneous pool of cells carrying targeted gene disruption across the population, circumventing clonal artifacts and providing a robust model for investigating L1CAM-dependent biology in a cancer-relevant context. The polyclonal nature reflects a realistic representation of the knockout phenotype without the selective pressures of single-cell cloning.
HT29 cells are epithelial cells originally isolated in 1964 from a primary colon adenocarcinoma of a 44-year-old Caucasian female. They are widely employed as a model for intestinal epithelial barrier function and colonic epithelial cell biology, retaining tumorigenic potential and presenting a well-characterized system for colorectal cancer research. Under defined culture conditions, HT29 cells can exhibit differentiated features, making them suitable for studying both undifferentiated and enterocytic phenotypes.
L1CAM is a transmembrane cell adhesion molecule of the immunoglobulin superfamily that mediates homophilic and heterophilic interactions critical for cell adhesion, migration, axon guidance, and neurite outgrowth. In cancer, L1CAM overexpression promotes invasiveness and metastasis by engaging integrin heterodimers such as ??5??1 and ??v??3, neuropilin-1, and fibroblast growth factor receptor (FGFR), leading to downstream activation of focal adhesion kinase (FAK), ERK1/2, and AKT. Upstream regulators include NF-??B, ??-catenin/TCF, TGF-??, SLUG (SNAI2), and SOX2, while downstream effectors comprise matrix metalloproteinases MMP-2 and MMP-9, which facilitate extracellular matrix degradation. L1CAM integrates signaling through MAPK/ERK (via MAP2K1, MAPK1, MAPK3), PI3K/AKT (PIK3CA, AKT1, AKT2), and NF-??B (NFKB1, RELA) cascades, with crosstalk to Wnt/??-catenin (CTNNB1, TCF4) and integrin-mediated adhesion (ITGB1, ITGA5) pathways, promoting epithelial-mesenchymal transition and aggressive tumor behavior.
In the HT29 colorectal cancer background, L1CAM knockout disables key adhesive and migratory programs, offering a precise model to dissect the molecular mechanisms driving colorectal cancer invasion and metastasis. This knockout system enables assessment of how loss of L1CAM alters integrin signaling, MAPK/ERK and AKT pathway activation, and EMT-associated gene expression. It serves as a valuable platform for evaluating L1CAM as a therapeutic target and for studying context-dependent signaling rewiring that may contribute to metastatic progression and drug resistance.
Researchers can utilize this polyclonal knockout population in a range of functional assays, including transwell migration, scratch wound healing, and cell adhesion assays to interrogate motility and attachment properties, as well as MTT and colony formation assays for proliferation, and annexin V apoptosis assays for cell death. Molecular profiling via western blotting, RT-qPCR, and immunofluorescence enables monitoring of key pathway components such as phosphorylated ERK1/2, AKT, and MMP expression. This model supports drug screening for anti-metastatic compounds and validation of L1CAM-associated biomarkers in colorectal and other metastatic cancers. For technical support and additional product information, please contact Ascent Research.
