The IGF2 Knockout T-47D Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population derived from the T-47D human breast carcinoma line, designed to disrupt the insulin-like growth factor 2 (IGF2) gene. This polyclonal pool provides a heterogeneous loss-of-function model, capturing varied editing events without clonal selection, suitable for population-level signaling and phenotypic analyses.
T-47D cells originate from a metastatic pleural effusion of an infiltrating ductal breast carcinoma in a 54-year-old female. They maintain a hormone-responsive phenotype, expressing estrogen receptor (ER), progesterone receptor (PR), and androgen receptor (AR), and are a benchmark model for studying luminal-type breast cancer and hormone-dependent growth regulation.
IGF2 encodes a secreted mitogenic peptide that binds IGF1R and insulin receptor (INSR) isoforms, leading to IRS1 phosphorylation and activation of the PI3K?CAKT and RAS?CMAPK cascades. AKT1 subsequently phosphorylates MTOR and RPS6KB1/S6K1 to promote protein synthesis, while parallel signaling through SHC, GRB2, and SOS triggers RAS?CRAF?CMEK?CERK, culminating in MAPK3/ERK1 nuclear translocation and CCND1 induction. IGF2 also suppresses apoptosis via AKT-dependent stabilization of BCL2. Signaling amplitude is modulated by the binding proteins IGFBP3 and the clearance receptor IGF2R, with upstream transcriptional control influenced by PLAG1, HMGA2, and the imprinted H19/IGF2 control region.
In T-47D cells, disruption of IGF2 is expected to dampen autocrine or paracrine mitogenic stimulation, leading to reduced phospho-AKT and phospho-ERK levels. This attenuation can manifest as decreased clonogenicity, slower proliferation, and impaired migration and invasion. The polyclonal nature ensures that phenotypes reflect population-average gene disruption rather than single-clone artifacts, providing a robust platform for mechanistic and pharmacological studies. The model is particularly relevant for understanding how growth factor signaling cooperates with hormone receptor activation in ER+ breast cancer.
These polyclonal knockout cells are suited for cancer biology, growth factor signaling, and drug target validation studies. Representative assays include western blotting for AKT and ERK phosphorylation, RT-qPCR for IGF2 transcript knockdown, MTT and BrdU proliferation assays, colony formation and transwell migration/invasion assays, flow cytometric cell cycle analysis, RNA-seq for transcriptomic profiling, and ELISA for secreted IGF2. Co-immunoprecipitation of IGF1R?CIRS1 complexes can further clarify altered signaling assemblies. For more information or to discuss specific experimental needs, please contact Ascent Research.
