This product is a CRISPR/Cas9-generated polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. The KCTD15 gene has been disrupted across the population, yielding a heterogeneous mixture of cells with diverse loss-of-function alleles. This format avoids clonal artifacts and provides a robust system for studying KCTD15-dependent processes. It is ideal for functional genomics and signaling pathway analysis in a colorectal cancer context.
HT29 cells originate from a primary colorectal adenocarcinoma of a 44-year-old female and exhibit epithelial morphology. They are widely used as a model for intestinal epithelial biology, drug absorption, and colorectal tumorigenesis. These cells harbor APC, p53, and BRAF mutations, leading to constitutive Wnt/??-catenin pathway activation and rapid proliferation, offering a relevant genetic backdrop for investigating KCTD15’s regulatory roles.
KCTD15 functions as a substrate adaptor for the CUL3-based E3 ubiquitin ligase, promoting ubiquitination and degradation of the GLI1 and GLI2 transcription factors to inhibit Hedgehog signaling. Additionally, KCTD15 interacts with ??-catenin and TCF/LEF complexes to modulate Wnt target gene expression. Upstream, KCTD15 is induced by BMP and Wnt signals, and it represses Hedgehog-responsive genes. Downstream targets include MYC and CCND1, linking KCTD15 to cell cycle control. Interactors such as SUFU, ZNF503, and TRIM32 further integrate these pathways.
In HT29 cells, where Wnt signaling is constitutively active due to APC truncations, KCTD15 disruption enables dissection of cross-talk between Hedgehog and Wnt pathways. Loss of KCTD15 de-represses GLI proteins and may alter Wnt target expression, affecting proliferation, differentiation, and tumorigenesis. This model is valuable for colorectal cancer research and for exploring KCTD15’s role in obesity-related metabolic signaling, given its genetic associations with adiposity.
Researchers can validate KCTD15 knockout by Sanger sequencing and western blot, and assess transcript levels via RT-qPCR. Functional assays include MTT, colony formation, and wound healing. Signaling output can be measured by TOP/FOP luciferase reporter for Wnt and dual-luciferase for Hedgehog. Co-immunoprecipitation identifies protein interactions, and RNA-seq enables transcriptomic profiling. For more information or custom inquiries, contact Ascent Research.
