The CCNY Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited human polyclonal knockout cell population derived from the 786-O clear cell renal cell carcinoma line. This product features targeted disruption of the CCNY gene, which encodes cyclin Y, a regulatory cyclin that activates cyclin-dependent kinases. The polyclonal nature reflects a heterogeneous pool of gene-edited cells, providing a robust loss-of-function model without the biases introduced by single-cell cloning. By disrupting CCNY gene function, this system enables functional interrogation of CCNY-dependent pathways in a disease-relevant context.
The parental 786-O cell line is derived from a primary clear cell renal cell carcinoma and carries a mutant VHL gene, recapitulating the most frequent genetic lesion in human RCC. This VHL deficiency leads to constitutive stabilization of hypoxia-inducible factors (HIFs) and a pseudo-hypoxic phenotype, rendering the cells highly dependent on alternative oncogenic drivers, such as Wnt signaling, for sustained proliferation. The 786-O background thus offers a pathophysiologically relevant model in which to investigate the tumor-promoting roles of CCNY.
CCNY encodes cyclin Y, which forms active complexes with CDK14 and CDK16 to phosphorylate the Wnt co-receptor LRP6 at key serine residues. This phosphorylation event triggers LRP6 signalosome assembly and downstream stabilization of ??-catenin (CTNNB1), allowing its nuclear translocation and transcriptional activation of MYC and CCND1. CCNY expression is positively regulated by the transcription factor E2F1 and by canonical Wnt ligand WNT3A, embedding it within a feed-forward loop that amplifies Wnt/??-catenin transduction. The cyclin Y?CCDK14/16 axis drives cell cycle progression, positioning CCNY at the intersection of cell cycle control and Wnt signaling.
In the context of clear cell renal cell carcinoma, CCNY-mediated signaling may synergize with VHL loss to fuel tumorigenic processes including uncontrolled proliferation and enhanced migratory capacity. Disrupting CCNY in 786-O cells allows researchers to dissect the contribution of cyclin Y to RCC biology independent of or in concert with hypoxic signaling. This knockout model can be used to evaluate the requirement for cyclin Y in maintaining oncogenic phenotypes, explore synthetic-lethal interactions with VHL deficiency, and identify potential therapeutic targets within the cyclin Y?CCDK14/16?CLRP6 cascade.
Researchers can utilize this CCNY knockout polyclonal population in a broad array of experimental assays. Wnt pathway activity can be assessed by TOPFlash dual-luciferase reporter systems or by monitoring ??-catenin levels via western blotting and RT-qPCR. Cell cycle distribution is readily measured by propidium iodide staining and flow cytometry, while proliferation and migration potentials are evaluated using standard in vitro assays. Transcriptional consequences of CCNY disruption can be profiled by RNA-seq, revealing downstream gene expression changes. The model is also suitable for drug sensitivity screening against cyclin-dependent kinase inhibitors or Wnt pathway antagonists. For additional information, please contact Ascent Research.
