The CCNY Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma line. The product comprises a heterogeneous pool of cells harboring target-gene disruption of CCNY, enabling robust functional loss-of-function studies without clonal selection biases.
The parental HCT 116 line is a human colorectal carcinoma epithelial cell line from a male patient, characterized by microsatellite instability-high (MSI-H) due to MLH1 deficiency, and oncogenic mutations in KRAS (G13D) and TP53, with a near-diploid karyotype. These genetic features establish HCT 116 as a key model for Wnt-driven colorectal cancer research.
CCNY encodes cyclin Y, a regulatory subunit that partners with CDK14 to phosphorylate the Wnt co-receptor LRP6 at Thr1472, potentiating Wnt/??-catenin signal transduction. This phosphorylation enhances downstream transcription of target genes including MYC, CCND1, and AXIN2 via the ??-catenin/TCF complex. The CCNY-CDK14 complex also promotes G2/M cell cycle progression, establishing a link between Wnt signaling and mitotic entry. CCNY expression is regulated by the Wnt/??-catenin/TCF transcriptional program and E2F1, and the cyclin Y-CDK14 complex interacts with destruction complex components APC and AXIN1. Canonical pathway components include Wnt ligands such as Wnt3a, Frizzled receptors, LRP6, Dishevelled (DVL), the destruction complex (AXIN1, APC, GSK3??), ??-catenin, and TCF4.
In HCT 116 cells, which harbor constitutive Wnt pathway activation, disruption of CCNY permits dissection of cyclin Y??s role in modulating Wnt output and cell cycle progression. Knockout of CCNY may reduce LRP6 phosphorylation and attenuate transcriptional activity of Wnt targets, influencing proliferation, colony formation, and tumorigenic potential. The model is particularly valuable for exploring how Wnt, KRAS, and cell cycle signals intersect in colorectal cancer, and for evaluating CCNY as a potential therapeutic target in Wnt-addicted malignancies such as colorectal, breast, hepatocellular, and glioma.
Researchers can utilize these polyclonal knockout cells in diverse applications. CCNY protein loss is validated by western blotting, and functional assessment includes TOP/FOP Flash reporter assays, RT-qPCR for MYC and AXIN2, and cell cycle analysis by propidium iodide staining. Co-immunoprecipitation confirms CCNY-CDK14 complex formation, while immunofluorescence monitors ??-catenin localization. Colony formation assays and drug sensitivity testing with Wnt inhibitors further probe tumorigenic phenotypes. For additional information, contact Ascent Research.
