The CCNY Knockout HEK293T Polyclonal Cells product consists of a population of HEK293T cells that have been edited using CRISPR/Cas9 to disrupt the CCNY gene, which encodes Cyclin Y. This polyclonal knockout cell population provides a loss-of-function model for studying the biological roles of Cyclin Y in a human cell context. The gene disruption is mediated by CRISPR/Cas9, generating a heterogeneous pool of edited alleles, and is suitable for pooled functional assays without clonal selection.
The host cell line HEK293T is a human embryonic kidney epithelial cell line widely used in biomedical research. These cells are transformed with adenovirus 5 DNA and stably express the SV40 large T antigen, which enables episomal replication of plasmids containing the SV40 origin of replication. HEK293T cells are highly transfectable and are frequently employed for transient protein expression, lentiviral production, and various cell-based assays, making them an ideal platform for generating knockout models to study gene function.
CCNY encodes Cyclin Y, a cyclin that partners with CDK14 to regulate Wnt/??-catenin signaling. Binding of Wnt ligands such as Wnt3a to Frizzled receptors triggers the Cyclin Y?CCDK14 complex to phosphorylate the co-receptor LRP6 on serine residues, a key event that stabilizes ??-catenin. Stabilized ??-catenin translocates to the nucleus, where it associates with TCF/LEF transcription factors to drive expression of proliferative genes including MYC and CCND1. Cyclin Y also interacts with DVL2, placing it within the wider Wnt signalosome. Through these actions, Cyclin Y promotes cell proliferation and migration, with additional links to PI3K/AKT and MAPK/ERK cascades.
In the HEK293T epithelial cell background, CCNY knockout provides a model to dissect Wnt/??-catenin signaling. Given Cyclin Y??s role in proliferation and migration, this knockout population enables assessment of these processes in a controlled setting. The polyclonal nature avoids clonal variation artifacts, and HEK293T??s high transfectability facilitates downstream molecular analyses. This model is relevant for studying signaling pathways in colorectal, lung, and breast cancers, as well as neurodegenerative diseases.
Typical applications include Western blot to monitor ??-catenin and phospho-LRP6 levels, RT-qPCR for Wnt target genes (e.g., AXIN2), and TOP/FOPFlash reporter assays to measure ??-catenin/TCF activity. Co-immunoprecipitation can assess Cyclin Y?CCDK14 binding, while proliferation (MTS/BrdU) and wound healing assays evaluate functional consequences of CCNY loss. Colony formation assays further assess clonogenic growth. This product serves cancer biology, signal transduction, and drug discovery research. Contact Ascent Research for technical details.
