The GSK3A Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line, featuring targeted disruption of the GSK3A gene. This heterogeneous pool of edited cells enables robust loss-of-function studies without clonal bias, suitable for investigating GSK3A-dependent signaling in cancer biology.
HCT 116 is a well-characterized epithelial colorectal carcinoma cell line with mismatch repair deficiency due to an MSH2 mutation and a KRAS G13D activating mutation, making it a standard model for tumorigenesis and genetic instability research. This genetic background provides a clinically relevant context for assessing the role of GSK3A in colorectal cancer.
GSK3A encodes a constitutively active serine/threonine kinase that phosphorylates ??-catenin, targeting it for ubiquitination and proteasomal degradation, thus acting as a negative regulator of the Wnt/??-catenin pathway. Kinase activity is inhibited by AKT-mediated phosphorylation at Ser21 in response to PI3K/AKT signaling and by Wnt ligand-induced disruption of the destruction complex composed of AXIN, APC, and DVL. Additionally, GSK3A phosphorylates downstream substrates such as glycogen synthase, c-Myc, cyclin D1, MCL1, and tau, integrating inputs from insulin, Hedgehog, and cell cycle pathways.
In HCT 116 cells, loss of GSK3A function can exacerbate ??-catenin stabilization and TCF/LEF transcriptional activity, potentially cooperating with oncogenic KRAS to drive proliferation and survival. This polyclonal knockout model is therefore instrumental for dissecting the crosstalk between Wnt and other mitogenic pathways, studying drug resistance mechanisms, and evaluating metabolic reprogramming in a colorectal cancer context.
The GSK3A knockout polyclonal cells are ideal for TOP/FOP flash reporter assays to gauge Wnt activity, western blotting for phospho-GSK3A or ??-catenin, MTS/MTT proliferation assays, colony formation, and Annexin V apoptosis detection. Additional applications include qRT-PCR for target genes (c-Myc, cyclin D1), co-immunoprecipitation of GSK3A with AXIN/??-catenin, and immunofluorescence to monitor ??-catenin localization. For further inquiries, contact Ascent Research.
