The BCL9 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited population of HT29 human colorectal adenocarcinoma cells featuring disruption of the BCL9 gene. This polyclonal knockout pool serves as a heterogeneous loss-of-function model for probing BCL9-dependent signaling and gene regulation without clonal selection bias.
The HT29 parental line is an adherent epithelial colorectal adenocarcinoma model from a 44-year-old female, harboring oncogenic mutations in TP53 (R273H), APC (E853* and T1556fs*3), and BRAF (V600E). These alterations lead to constitutive Wnt/??-catenin signaling via APC truncation and sustained MAPK pathway activity, making HT29 a relevant platform for colorectal cancer research.
BCL9 acts as a transcriptional coactivator bridging ??-catenin to PYGO1/PYGO2 and TCF/LEF transcription factors (e.g., TCF4, LEF1) at Wnt-responsive promoters. This scaffold facilitates recruitment of CBP/p300 and potentiates expression of target genes such as MYC, CCND1, AXIN2, and LGR5. Upstream Wnt3a stimulation engages Frizzled/LRP receptors, inhibits GSK-3??, and stabilizes ??-catenin, which accumulates in the nucleus and associates with BCL9 to drive pro-proliferative transcriptional programs.
In HT29 cells, where APC mutations render ??-catenin constitutively active, BCL9 is positioned as a critical mediator of oncogenic Wnt output. Its disruption is anticipated to impair ??-catenin complex assembly, attenuate MYC and CCND1 expression, and reduce proliferative capacity, offering a model to interrogate BCL9 dependency and Wnt-driven transcription in the context of BRAF and p53 mutant colorectal cancer.
These knockout cells are appropriate for functional assays including proliferation, colony formation, and migration/invasion studies. Transcriptional activity can be assessed via TOPFlash reporter, RT-qPCR for MYC/CCND1/AXIN2, and ChIP-qPCR for TCF/LEF binding. Co-immunoprecipitation can confirm disrupted ??-catenin?CBCL9 interactions. The model supports drug target validation, RNA-seq profiling, and pathway dissection. For additional information, please contact Ascent Research.
