The AXL Knockout HT29 Polyclonal Cells product comprises a heterogeneous population of HT29 human colorectal adenocarcinoma cells in which the AXL gene has been disrupted using CRISPR/Cas9 genome editing. This polyclonal knockout cell pool provides a versatile loss-of-function model for dissecting AXL-dependent biological functions without the clonal variability associated with single-cell-derived lines. By targeting the endogenous locus, the edited population enables robust and reproducible studies of AXL signaling in a cancer-relevant cellular background.
HT29 cells are derived from a 44-year-old female patient with colorectal adenocarcinoma and serve as a well-established model of intestinal epithelial biology. They exhibit epithelial morphology and retain many characteristics of colorectal tumors, including proficient DNA mismatch repair and expression of adenomatous polyposis coli (APC) proteins. Their widespread use in cancer research makes them a reliable platform for investigating tumor cell behavior, signaling networks, and therapeutic responses in colorectal cancer.
AXL encodes a receptor tyrosine kinase that transduces signals from the ligands Gas6 and Protein S, activating downstream pathways including PI3K/AKT and MAPK/ERK cascades. In the HT29 context, AXL engages adaptor proteins such as Grb2 and PI3K and associates with regulators like SHP2, SOCS1, and CD44. Activation leads to phosphorylation of AKT and ERK, promoting cell survival, proliferation, and migration. AXL signaling also upregulates transcription factors SNAI1 and SLUG, driving epithelial-mesenchymal transition (EMT), and enhances expression of matrix metalloproteinase MMP9 and immune checkpoint molecule PD-L1.
Disruption of AXL in HT29 polyclonal cells dampens Gas6-mediated activation of PI3K/AKT and MAPK/ERK pathways, resulting in reduced phosphorylation of AKT and ERK and attenuated downstream transcriptional programs. This diminishes cell viability, proliferative capacity, and migratory/invasive potential. Furthermore, loss of AXL impairs EMT induction and lowers PD-L1 levels, thereby weakening immune evasion mechanisms. These effects highlight the critical role of AXL in maintaining colorectal adenocarcinoma cell aggressiveness and therapy resistance.
These polyclonal knockout cells are suitable for a wide array of research applications, including the study of colorectal cancer biology, drug resistance, metastasis, and immune checkpoint regulation. Researchers can employ them in mechanistic investigations of TAM receptor signaling, EMT, and PI3K/AKT and MAPK/ERK pathway modulation. Representative assays include Western blotting for total and phospho-AKT/ERK, RT-qPCR for AXL and its downstream targets (e.g., SNAI1, SLUG, PD-L1), flow cytometry for apoptosis and proliferation markers, Transwell migration/invasion assays, and drug sensitivity testing. For additional technical details, please contact Ascent Research.
