The DIP2A Knockout 786-O Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population in which the DIP2A gene has been disrupted in the 786-O human cell line. This polyclonal pool offers a heterogeneous population of cells with targeted gene disruption, enabling loss-of-function studies of DIP2A in a well-characterized renal carcinoma background. The knockout model is generated using CRISPR/Cas9-mediated gene editing to introduce disruptive modifications within the DIP2A locus, resulting in a polyclonal population suitable for functional assays without the need for clonal isolation.
The 786-O cell line is derived from a primary clear cell renal cell carcinoma (ccRCC), representing malignant kidney epithelial cells. As a widely used in vitro model for renal cell carcinoma, 786-O cells exhibit features characteristic of ccRCC, including dysregulated hypoxia signaling and constitutive activation of downstream growth and survival pathways. This genetic context makes 786-O cells an appropriate host for investigating oncogenic mechanisms, particularly those involving PI3K/AKT/mTOR signaling and metabolic reprogramming.
DIP2A encodes a transmembrane receptor that binds the secreted ligand FSTL1, thereby initiating intracellular signaling cascades. Upon FSTL1 stimulation, DIP2A activates the PI3K/AKT/mTOR pathway, leading to phosphorylation of AKT1, mTOR, and its downstream effectors S6K and 4E-BP1. DIP2A also influences acetyl-CoA metabolism through regulation of acetyl-CoA carboxylase. Additionally, DIP2A has been reported to interact with DMAP1 and DNMT1, suggesting roles in transcriptional and epigenetic regulation. In the context of neural development, DIP2A participates in axon guidance, and genetic variants are linked to developmental dyslexia. The knockout of DIP2A in 786-O cells is expected to abrogate FSTL1-induced AKT phosphorylation and may perturb acetyl-CoA metabolic flux.
In 786-O ccRCC cells, the FSTL1/DIP2A signaling axis is hypothesized to promote cell proliferation and survival through sustained activation of the PI3K/AKT/mTOR cascade. HIF1A, a transcription factor frequently stabilized in ccRCC due to VHL inactivation, may serve as an upstream regulator that upregulates DIP2A expression under hypoxic conditions. By disrupting DIP2A, this polyclonal knockout model enables researchers to dissect the contribution of DIP2A-mediated signaling to kidney cancer progression, including its impact on cell growth, migration, invasion, and metabolic responses. The model is particularly valuable for dissecting how FSTL1/DIP2A signals intersect with oncogenic drivers in ccRCC.
Typical applications include western blot analysis of AKT/mTOR pathway activation, cell viability assays (MTT or CellTiter-Glo), Transwell migration/invasion assays, acetyl-CoA quantification, and transcriptomic profiling by RNA-seq to analyze gene expression changes upon DIP2A loss. Co-immunoprecipitation experiments can be performed to confirm disrupted FSTL1-DIP2A interaction. The polyclonal knockout population is suitable for both short-term signaling and long-term functional studies, offering a convenient tool to investigate DIP2A as a potential therapeutic target in renal cell carcinoma. For further technical specifications or support, please contact Ascent Research.
