The KDM5D Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited heterogeneous population with targeted disruption of KDM5D, designed for loss-of-function analysis of this Y-chromosome histone demethylase. As polyclonal knockout cells, they offer a population-level model to study epigenetic regulation without clonal selection biases. This product enables investigation of KDM5D-dependent transcriptional repression mechanisms in a well-characterized human embryonic kidney epithelial host.
HEK293T cells derive from HEK293, originally established from human embryonic kidney, and are transformed with adenovirus 5 DNA. They constitutively express SV40 large T antigen, facilitating episomal plasmid replication and high-level transient protein expression. Renowned for efficient viral packaging and stable gene expression, HEK293T serves as a robust platform for gene editing studies, offering rapid growth and high transfectability ideal for functional genomics.
KDM5D specifically demethylates histone H3 lysine 4 trimethylation (H3K4me3) and dimethylation (H3K4me2) to the monomethylated form, acting as a transcriptional repressor. It functions within large complexes containing HDAC1, HDAC2, SIN3A, REST, and RbBP5, linking demethylation to deacetylation. Upstream factors such as androgen receptor and SOX9 regulate KDM5D, while downstream targets include CCND1, CDKN1A, and TP53, positioning it at the nexus of chromatin remodeling, cell cycle control, and DNA damage response. This demethylase is critical for spermatogenesis and male-specific gene regulation, with its dysfunction linked to testicular cancer, prostate cancer, and colorectal cancer.
In HEK293T, which retains Y chromosome gene expression, KDM5D knockout likely elevates H3K4me3 levels at target promoters, upregulating CCND1 and CDKN1A and potentially altering proliferation dynamics. The polyclonal knockout model captures a spectrum of editing events, mimicking heterogeneous loss-of-function scenarios and enabling studies of KDM5D??s role in androgen receptor-driven pathways and epigenetic reprogramming relevant to Y chromosome loss in aging and malignancy.
Researchers can employ this product for ChIP-qPCR to quantify H3K4me3 enrichment at gene promoters, RT-qPCR and western blotting to measure target gene and protein changes, and immunofluorescence to assess global chromatin modifications. Cell cycle profiling by flow cytometry, proliferation assays, and RNA-seq transcriptomics further illuminate KDM5D??s impact. The knockout cells also facilitate histone demethylase inhibitor screening and functional investigation of Y chromosome genes in cancer. For technical inquiries, contact Ascent Research.
