The HDAC8 knockout HT29 polyclonal cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 human colorectal adenocarcinoma cell line, designed for HDAC8 loss-of-function studies. This mixed cell population carries targeted gene disruptions introduced by CRISPR/Cas9 editing, enabling robust analyses without clonal selection artifacts.
HT29 cells are a widely used in vitro model for colorectal carcinoma, isolated from a 44-year-old female patient with colorectal adenocarcinoma. These adherent epithelial cells exhibit a well-differentiated phenotype, carry a mutant TP53 gene, and are microsatellite stable. They serve as a relevant system to study intestinal epithelial barrier function, drug response, and oncogenic signaling pathways in colorectal cancer.
HDAC8 is a class I histone deacetylase that removes acetyl groups from lysine residues on histones (H3, H4) and non-histone substrates such as the cohesin subunit SMC3 and the tumor suppressor p53 (TP53). Deacetylation of SMC3 by HDAC8 is essential for cohesin complex recycling during the cell cycle, while p53 deacetylation modulates its transcriptional activity and protein stability. HDAC8 expression and activity are regulated by miR-15a, miR-16, cAMP/PKA signaling, CREB, and SP1. The enzyme interacts with corepressors NCOR1 and SMRT, and forms functional complexes with RAD21 and MEF2C. Downstream, HDAC8 influences acetylation of histones H3/H4, and modulates ERR?? and Cortactin, linking its deacetylase function to chromatin remodeling, metabolism, and cytoskeletal dynamics.
In HT29 colorectal cancer cells, HDAC8 knockout disrupts the balance of acetylation on histones and non-histone substrates, leading to impaired cell cycle progression and apoptosis. Loss of HDAC8 function destabilizes cohesin complex dynamics, alters p53 activity, and modifies histone acetylation marks such as H3K9ac and H4K16ac. This polyclonal knockout population provides a relevant model to dissect HDAC8-dependent pathways in a colorectal cancer background, with potential relevance to Cornelia de Lange syndrome, neuroblastoma, and T-cell acute lymphoblastic leukemia research.
Researchers can utilize these cells in various assays to investigate HDAC8 biology and screen therapeutic compounds. Typical applications include Western blotting for HDAC8, acetyl-SMC3, and acetyl-p53; RT-qPCR for downstream transcriptional changes; ChIP-qPCR for H3K9ac and H4K16ac; co-immunoprecipitation of HDAC8?CSMC3 complexes; flow cytometric cell cycle analysis; and Annexin V apoptosis assays. These polyclonal knockout cells are ideal for cancer epigenetics research, HDAC inhibitor testing, cohesin complex studies, and colorectal cancer drug response investigations. For detailed technical specifications or ordering information, please contact Ascent Research.
