The KCTD9 Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This heterogeneous pool carries targeted disruption of the KCTD9 gene, creating a loss-of-function model suitable for population-level functional studies without clonal expansion. The polyclonal format ensures a cost-effective, ready-to-use system for investigating KCTD9 biological functions in colorectal cancer.
HT29 is an epithelial colorectal adenocarcinoma cell line isolated from a 44-year-old female, widely used as a colorectal cancer model. These cells carry a BRAF-V600E mutation and an APC mutation that drives constitutive Wnt/??-catenin signaling, and they are capable of enterocytic differentiation. The line also expresses functional NF-??B pathway components, making it suitable for dissecting signaling crosstalk in a defined genetic context.
KCTD9 functions as a substrate-specific adaptor for the CUL3-RING E3 ubiquitin ligase complex, interacting with CUL3 and RBX1 to promote ubiquitination and proteasomal degradation of target proteins, including TNFAIP3 (A20), HDAC1, and ??-catenin (CTNNB1). KCTD9 expression is regulated by TP53, DNA damage signals, and MYC. Upon KCTD9 knockout, accumulation of these substrates occurs, with TNFAIP3 stabilization predicted to modulate NF-??B activity, ??-catenin accumulation enhancing TCF/LEF-driven transcription, and HDAC1 potentially altering chromatin remodeling. Thus, KCTD9 disruption reconfigures both NF-??B and Wnt/??-catenin signaling nodes.
In HT29 cells, which harbor intrinsic APC-mutation-driven Wnt pathway activation, loss of KCTD9 is expected to further augment ??-catenin-dependent transcription and enhance NF-??B signaling, potentially promoting proliferation and survival. This model thus enables investigation of KCTD9??s tumor-suppressive roles and the interplay between ubiquitin-mediated proteolysis and oncogenic signaling. It provides a defined system to assess how KCTD9 loss impacts colorectal cancer cell behavior.
This product is designed for functional assays including western blotting and RT-qPCR for KCTD9 and substrate analysis, NF-??B and TCF/LEF luciferase reporters, cell proliferation and apoptosis measurements, co-immunoprecipitation with CUL3, and ubiquitination assays. The cells are also valuable for drug screening and genomic studies investigating ubiquitin-proteasome targeting. For technical inquiries or custom projects, please contact Ascent Research.
