The KREMEN2 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the human KREMEN2 gene has been disrupted to eliminate functional KREMEN2 protein expression. This heterogeneous pool of knockout cells provides a robust loss-of-function model for studying the regulatory roles of KREMEN2 in Wnt/??-catenin signal transduction. The polyclonal format ensures genetic diversity while maintaining consistent target-gene ablation, making it suitable for high-throughput screening and pathway analysis.
The parental HEK293T cell line is a widely utilized human embryonic kidney epithelial model that stably expresses the SV40 large T antigen. This feature enables episomal replication of plasmids containing the SV40 origin of replication, yielding high transfection efficiency and robust exogenous protein expression. HEK293T cells are routinely employed for recombinant protein production, lentiviral packaging, and transient reporter assays, providing a versatile platform for interrogating signal transduction mechanisms.
KREMEN2 encodes a type I transmembrane protein that functions as a high-affinity co-receptor for Dickkopf proteins (DKK1, DKK2, and DKK4). Upon ligand binding, KREMEN2 forms a ternary complex with the Wnt co-receptor LRP6, triggering clathrin-mediated endocytosis and lysosomal degradation of LRP6. This process potently inhibits the canonical Wnt/??-catenin pathway. Downstream consequences include reduced stabilization and nuclear accumulation of ??-catenin, leading to diminished transcription of Wnt target genes such as MYC, CCND1, AXIN2, and LEF1. The pathway involves key components including WNT3A, Frizzled receptors, Dishevelled (DVL), AXIN, APC, GSK3??, and TCF/LEF transcription factors.
In the HEK293T background, disruption of KREMEN2 relieves the negative regulation exerted on LRP6, thereby augmenting Wnt/??-catenin signaling output. This model is particularly informative for dissecting the interplay between DKK-mediated inhibition and receptor availability, as HEK293T cells possess intact Wnt pathway machinery and are amenable to cotransfection with pathway reporters and expression constructs. The polyclonal nature reduces artefacts arising from clonal selection, enabling physiologically relevant studies of signaling dynamics and feedback regulation.
The KREMEN2 Knockout HEK293T Polyclonal Cells are ideally suited for a range of experimental applications, including TOP/FOP Flash luciferase reporter assays to quantify ??-catenin-dependent transcription, Western blotting for phospho-LRP6 and total ??-catenin levels, and co-immunoprecipitation to assess DKK1?CKREMEN2 interactions. Further uses encompass RT-qPCR profiling of Wnt target genes (AXIN2, MYC), immunofluorescence analysis of ??-catenin subcellular localization, and functional assays measuring cell proliferation and migration. For additional technical information, protocols, or ordering details, please contact Ascent Research.
