The KDM6A Knockout HT29 Polyclonal Cells are a heterogeneous population of HT29 human colorectal adenocarcinoma cells edited by CRISPR/Cas9 to disrupt the KDM6A gene. This polyclonal knockout pool, generated without clonal selection, provides a loss-of-function model for studying KDM6A-dependent epigenetic regulation while minimizing clonal artifacts commonly associated with single-cell derived lines.
HT29 is an adherent epithelial cell line established from a primary colon adenocarcinoma of a 44-year-old female. It harbors oncogenic mutations in APC, TP53, and KRAS, and retains the capacity to differentiate into enterocyte-like cells under defined conditions, making it a valuable model for colorectal cancer biology and intestinal epithelial transport studies.
KDM6A (UTX) functions as a histone H3K27 demethylase that removes repressive H3K27me2/me3 marks, leading to transcriptional activation. It forms complexes with MLL3/MLL4 (KMT2C/KMT2D), ASH2L, RBBP5, and WDR5, and is regulated by TGF-?? and retinoic acid signaling. Upon activation, KDM6A interacts with SMAD2/3 and RB1 to modulate key targets such as CDKN1A, CDH1, and WNT pathway inhibitors, thereby intersecting with Wnt/??-catenin (CTNNB1/TCF7L2), Notch, and retinoblastoma pathways to maintain a tumor-suppressive chromatin state.
In HT29 cells, loss of KDM6A activity elevates H3K27me3 levels, leading to silencing of tumor suppressors and aberrant activation of developmental pathways. Coupled with the cell line??s existing APC and KRAS mutations, KDM6A knockout promotes dedifferentiation, enhances proliferation, and facilitates invasive behavior, providing a relevant model to study epigenetic dysregulation in colorectal cancer progression and to explore cooperative mechanisms between genetic and epigenetic lesions.
These polyclonal knockout cells are suitable for epigenetic profiling by western blot and immunofluorescence for H3K27me3, gene expression analysis by RT-qPCR and RNA?seq, and chromatin immunoprecipitation (ChIP?qPCR) to assess histone modifications at specific promoters. Functional studies may include cell proliferation, colony formation, migration/invasion, and apoptosis assays to evaluate tumor cell properties. The model also enables screening of histone demethylase inhibitors and investigation of tumor?Cmicroenvironment interactions. For additional technical details or custom requests, please contact Ascent Research.
