The IGF2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, featuring targeted disruption of the insulin-like growth factor 2 (IGF2) gene. As a polyclonal population, this product comprises a heterogeneous mix of edited alleles, providing a robust loss-of-function model that avoids clonal selection artifacts and ensures reproducible functional studies. The CRISPR/Cas9-mediated gene disruption eliminates functional IGF2 protein expression, thereby abrogating the autocrine and paracrine signaling cascades normally initiated by this growth factor.
HT29 cells are a well-characterized human colorectal adenocarcinoma epithelial line extensively used in cancer biology and drug discovery. Originating from a primary colon adenocarcinoma, these cells grow adherently and exhibit moderate differentiation, expressing key surface receptors involved in cell proliferation and survival. Their colorectal origin and well-documented molecular landscape make HT29 an ideal host for knockout studies focused on signaling pathways dysregulated in colon cancer, particularly those driven by growth factor ligands.
IGF2 is a pivotal fetal mitogen that exerts its effects primarily through binding to the insulin-like growth factor 1 receptor (IGF1R) and the insulin receptor (INSR). Ligand engagement triggers receptor autophosphorylation and recruitment of adaptor proteins such as IRS1, SHC, and GRB2, leading to activation of the PI3K-AKT and MAPK-ERK signaling modules. Key downstream effectors include AKT1, mTOR, S6 kinase, and ERK1/2, which collectively promote cell cycle progression, protein synthesis, and inhibition of apoptosis. IGF2 bioavailability is tightly regulated by a family of six IGF-binding proteins (IGFBP1-6) and by clearance through IGF2R. Additionally, its expression is controlled by an imprinting mechanism involving upstream regulators such as CTCF, the H19 long non-coding RNA, and PLAG1. In the knockout cells, loss of IGF2 protein prevents activation of these receptor-mediated pathways, leading to attenuation of proliferative and survival signals.
In the context of HT29 colorectal adenocarcinoma, autocrine IGF2 signaling significantly contributes to tumor cell growth, migration, and resistance to apoptotic stimuli. Disruption of IGF2 in this cellular background offers a powerful model to dissect the ligand’s specific contributions to malignant phenotypes. Because HT29 cells retain partial differentiation capacity, the knockout also enables exploration of how IGF2 loss influences epithelial-mesenchymal transition and sensitivity to conventional chemotherapeutics. Furthermore, given the imprinting status of IGF2 and its association with developmental disorders such as Beckwith-Wiedemann syndrome and Silver-Russell syndrome, this model facilitates epigenetic studies on imprinting dysregulation in cancer.
These polyclonal knockout cells are suited for a broad range of functional assays, including western blotting and RT-qPCR to validate target disruption and downstream signaling changes, cell proliferation (MTT/BrdU), colony formation, migration (scratch/transwell), and apoptosis (Annexin V/PI) analyses. Phospho-AKT ELISA can quantify pathway inhibition, and in vivo xenograft tumor growth assays assess tumorigenic potential. Applications span colorectal cancer research, growth factor signaling investigations, drug target validation (e.g., IGF1R inhibitors), and studies of genomic imprinting in cancer. For further information, please contact Ascent Research.
