The IGFBP5 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population generated from the HT29 human colorectal adenocarcinoma cell line, featuring targeted disruption of the IGFBP5 gene. This product provides a heterogeneous pool of cells with loss-of-function mutations in IGFBP5, enabling functional analysis of its role in colorectal cancer biology without the constraints of single-cell clonal selection. The polyclonal format better preserves the genetic diversity encountered in tumor populations, making it suitable for studying gene function in a more biologically relevant context.
The HT29 cell line was established from a 44-year-old female patient with colorectal adenocarcinoma and retains key epithelial characteristics, including the ability to form polarized monolayers and express intestinal differentiation markers. It is widely used as a model for intestinal epithelial biology, differentiation, barrier function, and colorectal cancer. This well-characterized line provides a robust platform for examining the impact of genetic modifications on signaling pathways and therapeutic responses in colorectal cancer research.
IGFBP5 encodes a carrier protein that binds insulin-like growth factors (IGF-I and IGF-II), modulating their half-life and receptor accessibility. It also exhibits IGF-independent functions by interacting with fibronectin, vitronectin, and integrins to influence proliferation, apoptosis, and migration. Upstream regulators such as p53, TGF-??, retinoic acid, FGFs, and EGF control its expression. Downstream, IGFBP5 affects AKT and ERK1/2 phosphorylation, and alters expression of p21, cyclin D1, Bcl-2, Bax, MMP-9, and E-cadherin. Knockout of IGFBP5 removes this regulatory layer, resulting in heightened IGF-mediated PI3K/AKT and MAPK/ERK signaling, thereby shifting the balance of cell growth, survival, and motility.
In the context of HT29 colorectal adenocarcinoma cells, IGFBP5 displays context-dependent roles, acting as either a tumor suppressor or promoter depending on microenvironmental cues and molecular context. The IGFBP5 knockout model enables systematic dissection of these dual functions. By eliminating IGFBP5, researchers can assess its impact on key oncogenic pathways, evaluate changes in epithelial-mesenchymal transition markers, and investigate how loss of IGFBP5 affects sensitivity to chemotherapeutic agents. This model is particularly valuable for exploring the interplay between IGF signaling and other pathways such as Wnt and TGF-??, which are frequently dysregulated in colorectal cancer.
Typical research applications include western blotting and phospho-signaling analysis to quantify pathway activation, RT-qPCR and RNA-seq for transcriptional profiling, immunofluorescence and flow cytometry to assess protein expression and cell cycle dynamics, and functional assays including apoptosis detection, migration/invasion chambers, and drug sensitivity studies. The polyclonal IGFBP5 knockout HT29 population supports investigations into the role of IGFBP5 in colorectal cancer progression, metastasis, and therapeutic resistance, offering a versatile platform for both basic and translational research. For additional information, please contact Ascent Research.
