The DIP2A Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma epithelial cell line. This product provides a heterogeneous pool of cells carrying targeted disruption of the DIP2A gene, enabling loss-of-function studies in a genetically defined colorectal cancer background. As a polyclonal population, these cells retain genetic diversity from independent editing events, facilitating pooled functional screens and bulk biochemical analyses.
The parental HCT 116 cell line is a widely used model of colorectal carcinoma, featuring an MLH1 mutation that leads to high microsatellite instability (MSI-high) and a KRAS G13D oncogenic driver mutation, while retaining wild-type p53. These adherent epithelial cells are tumorigenic and harbor activated AKT/mTOR signaling, making them a suitable host for studying DIP2A-mediated survival mechanisms and therapeutic responses in colorectal cancer.
DIP2A acts as a cell surface receptor for the secreted glycoprotein Follistatin-like 1 (FSTL1). Upon ligand binding, DIP2A recruits and activates AKT1, leading to downstream phosphorylation of mTOR and S6 kinase (S6K). This FSTL1?CDIP2A?CAKT1?CmTOR cascade upregulates anti-apoptotic proteins such as Bcl-2, promoting cell survival and proliferation while inhibiting caspase-mediated apoptosis. DIP2A thus serves as a critical mediator of FSTL1-dependent trophic signaling in colorectal epithelial cells.
In HCT 116 cells, DIP2A knockout disrupts FSTL1-mediated survival signaling, allowing dissection of its contribution to colorectal cancer proliferation and apoptosis resistance. The coexistence of KRAS G13D and DIP2A in this line enables interrogation of the interplay between oncogenic RAS signals and the FSTL1?CAKT1?CmTOR axis. Researchers can examine how loss of DIP2A affects downstream phosphorylation, cell cycle progression, and chemosensitivity, providing insights into pathway dependencies in MSI-high tumors.
Key applications involve Western blotting for phospho-AKT (Ser473), phospho-mTOR (Ser2448), and phospho-S6K (Thr389); cell proliferation assays; and caspase-3/7 apoptosis assays. Co-immunoprecipitation verifies DIP2A?CFSTL1 interaction, while RT-qPCR quantifies Bcl-2 transcript levels. The model is applicable to mTOR/AKT inhibitor screening, offering a platform to evaluate compound efficacy in modulating FSTL1-dependent survival pathways. For additional technical information, please contact Ascent Research.
