The IGF2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited cell population with disrupted IGF2 gene in the HAP1 near-haploid human cell line, providing a loss-of-function model for studying insulin-like growth factor 2 (IGF2) signaling. This polyclonal pool captures a range of editing outcomes, minimizing clonal artifacts and enabling robust population-level functional analyses. Homo sapiens IGF2 is a mitogenic peptide hormone implicated in proliferation and survival pathways, and this knockout tool supports rigorous investigation of its role in cancer and growth disorders.
HAP1 is a male-origin, adherent, fibroblast-like cell line derived from a chronic myeloid leukemia patient, retaining the BCR-ABL fusion oncogene. Its near-haploid karyotype ensures that a single allelic disruption can result in complete loss of gene function, facilitating clear genotype-phenotype associations. This cell line is widely adopted for genetic screening and knockout studies due to its stable growth and suitability for high-throughput assays, including proliferation, apoptosis, and migration analyses.
IGF2 functions primarily through IGF1 receptor (IGF1R) binding, activating the PI3K/AKT and MAPK/ERK cascades. Mechanistically, IGF2-IGF1R engagement phosphorylates IRS1, stimulating PI3K-mediated AKT1 activation that regulates mTOR and FOXO1-dependent survival and metabolism. Concurrently, SHC-GRB2-SOS signaling turns on RAS, which via RAF1, MAP2K1, and ERK1/2 (MAPK3/MAPK1) drives transcription of proliferation factors like MYC and CCND1 (cyclin D1). IGF2 expression is controlled by transcription factors PLAG1, SP1, and AP-2, and modulated by growth hormone and imprinting. Its bioavailability is regulated by six IGF-binding proteins (IGFBP1?C6) and clearance receptor IGF2R.
Disruption of IGF2 in HAP1 cells interrupts ligand delivery to IGF1R, thereby dampening AKT and ERK pathway activity, which are critical for cell growth and survival. This knockout model is valuable for dissecting how IGF2 loss interacts with the BCR-ABL-driven oncogenic program inherent to the HAP1 background. It provides a platform to study the pathological roles of IGF2 in Beckwith-Wiedemann syndrome, colorectal, breast, and hepatocellular carcinomas, and Wilms tumor, and to explore the effects of epigenetic imprinting alterations in a controlled genetic setting.
Research applications include functional genomics, drug target validation, and pathway inhibitor screening. Assays such as RT-qPCR for IGF2, immunoblotting for phospho-AKT1 and phospho-ERK1/2, MTS/WST-1 proliferation, Annexin V apoptosis, colony formation, and RNA-seq transcriptomics are routinely performed. These polyclonal knockout cells also facilitate receptor-ligand interaction studies and screening of small-molecule inhibitors targeting IGF1R or downstream kinases. For additional technical inquiries, please contact Ascent Research.
