The IGF2R Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line, carrying targeted disruption of the IGF2R gene. This polyclonal pool, generated without single-cell cloning, preserves heterogeneous genetic backgrounds while disrupting IGF2R expression through CRISPR/Cas9-mediated gene editing, offering a versatile loss-of-function model for studying IGF2R-dependent signaling and lysosomal trafficking in a colon cancer context.
HT29 is a well-established human colorectal adenocarcinoma epithelial cell line originally isolated from a 44-year-old female patient. These cells are widely employed as a model for colon cancer research, mucin secretion, and intestinal barrier function. Notably, HT29 cells display phenotypic plasticity, able to differentiate into enterocyte-like and goblet-like cells under appropriate culture conditions, making them particularly suitable for investigating colonic epithelial biology and tumor progression.
The IGF2R gene encodes a multifunctional type I transmembrane receptor that acts as a scavenger for insulin-like growth factor 2 (IGF2) and mannose-6-phosphate (M6P)-tagged lysosomal hydrolases. By binding and internalizing extracellular IGF2, IGF2R attenuates signaling through the IGF1R-AKT/ERK mitogenic pathway, thus restraining cell proliferation and survival. Concurrently, IGF2R mediates the delivery of newly synthesized M6P-modified lysosomal enzymes, such as cathepsins, to the lysosome, maintaining proteolytic homeostasis. Additionally, IGF2R interacts with latent TGF-beta 1 (TGFB1) complexes involving latency-associated peptide (LAP) and facilitates TGF-beta activation, linking the receptor to apoptosis and extracellular matrix regulation.
Disruption of IGF2R in HT29 cells is predicted to elevate extracellular IGF2 concentrations, thereby enhancing ligand availability for IGF1R and resulting in sustained activation of AKT and ERK signaling cascades. This pro-survival and proliferative shift aligns with the tumorigenic phenotype of colorectal cancer. Moreover, the loss of M6P-dependent lysosomal enzyme trafficking is expected to perturb lysosomal function and cathepsin distribution, potentially affecting autophagy, antigen presentation, and degradation pathways. The combined dysregulation of IGF and TGF-beta pathways in this knockout model provides a powerful platform to dissect the tumor-suppressive roles of IGF2R in colorectal adenocarcinoma.
This polyclonal knockout cell population is suited for a range of research applications, including investigations into colon cancer proliferation and metastasis, IGF2-driven tumorigenesis, lysosomal trafficking and metabolic control, and TGF-beta signaling crosstalk. Representative assays include western blotting for phospho-AKT and phospho-ERK, RT-qPCR for residual IGF2R transcript, MTT proliferation assays, Transwell invasion studies, caspase-3/7 apoptosis assays, and immunofluorescence for LAMP1 and cathepsins. The model also facilitates drug sensitivity and resistance testing in the context of enhanced IGF1R signaling. For additional technical details, please contact Ascent Research.
