The DVL3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with disruption of the DVL3 gene in the human HT29 colorectal adenocarcinoma cell line. This loss-of-function model eliminates DVL3 scaffold protein expression, enabling dissection of its role in Wnt signaling.
The HT29 cell line, derived from a primary colorectal adenocarcinoma of a Caucasian female, is a widely used model of intestinal epithelial biology. These adherent cells can differentiate into enterocyte-like cells, facilitating studies of intestinal differentiation, mucus production, and colorectal cancer. With aberrant Wnt/??-catenin signaling due to APC mutations, HT29 cells offer a relevant context for examining DVL3-dependent pathway regulation.
DVL3 encodes a cytoplasmic phosphoprotein that serves as a pivotal scaffold in Wnt signal transduction. Upon activation by Wnt ligands (Wnt3a, Wnt5a) and Frizzled receptors (FZD1-10), DVL3 is recruited to co-receptors LRP5/6 and polymerizes, a process regulated by CK1?? and CK2. This polymerization interacts with AXIN1/AXIN2, inhibiting the ??-catenin destruction complex (APC, GSK3B, CK1??), leading to ??-catenin stabilization and nuclear translocation, which activates target genes MYC, CCND1, and AXIN2 via TCF/LEF. DVL3 also directs non-canonical pathways, including JNK and RHO/ROCK signaling, influencing cell polarity and migration. Disruption of DVL3 therefore impairs both canonical and non-canonical Wnt branches.
In HT29 cells, DVL3 knockout provides a reductionist model for Wnt pathway dissection. Despite constitutive ??-catenin activity from APC loss, DVL3 ablation can further attenuate signaling, probing threshold requirements. Loss of DVL3 is expected to reduce proliferation via MYC and CCND1 downregulation and impair invasive properties driven by non-canonical effectors, making the model ideal for studying tumor cell growth and differentiation.
These polyclonal knockout cells support standard Wnt assays: Western blotting for ??-catenin and phospho-LRP6, TOPFlash/FOPFlash reporter assays for TCF/LEF activity, and RT-qPCR for MYC and AXIN2. Immunofluorescence tracks ??-catenin localization, while co-immunoprecipitation maps DVL3-AXIN interactions. Applications include drug screening for Wnt inhibitors and cell migration/invasion assays in colorectal cancer research. For further information, contact Ascent Research.
