The CCPG1 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human clear cell renal cell carcinoma line 786-O, designed to disrupt CCPG1 gene function. This polyclonal pool provides a heterogeneous loss-of-function model that minimizes clonal artifacts, enabling robust functional analyses of CCPG1 in a VHL-deficient, HIF-activated cellular context.
The parental 786-O cell line is a well-characterized human clear cell renal cell carcinoma (ccRCC) model, originally derived from a primary adenocarcinoma. These adherent epithelial cells carry a VHL mutation that disrupts pVHL function, resulting in constitutive stabilization of hypoxia-inducible factors (HIFs). This drives transcriptional programs that enhance angiogenesis, metabolic rewiring, and tumor progression, making 786-O an essential tool for investigating ccRCC biology, including mechanisms of metastasis and therapeutic resistance.
CCPG1 is a p53-inducible inhibitor of the G1/S cell cycle transition. Upon p53 activation, CCPG1 directly binds and inhibits CDK2, preventing CDK2-Cyclin E-mediated phosphorylation of Rb. Hypophosphorylated Rb sequesters E2F transcription factors, blocking entry into S phase. CCPG1 thus operates within the p53-p21-CDK2-Cyclin E-Rb-E2F signaling axis to enforce cell cycle arrest downstream of DNA damage signals.
In VHL-deficient 786-O cells, constitutive HIF signaling creates a unique context for examining the intersection of hypoxia responses and cell cycle control. Knocking out CCPG1 enables dissection of p53-dependent G1/S arrest under HIF activation and assessment of the CCPG1-CDK2 checkpoint dependency in ccRCC. This model is valuable for testing CDK inhibitors and exploring synthetic lethal vulnerabilities.
This knockout model supports diverse functional assays, including western blotting to assess CDK2, Cyclin E, p53, and p21 expression; RT-qPCR to quantify CCPG1 transcript levels; and flow cytometry for detailed cell cycle distribution analysis. Proliferation can be measured by MTT or BrdU incorporation, and co-immunoprecipitation can validate the CCPG1-CDK2 interaction. Additional applications include soft agar colony formation for anchorage-independent growth, Annexin V apoptosis assays, and migration/invasion studies. The polyclonal pool is particularly suited for drug sensitivity screening, especially with CDK inhibitors. For further information, please contact Ascent Research.
