The KDM4A Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of KDM4A. This product comprises a heterogeneous pool of HEK293T cells harboring targeted disruptions at the endogenous KDM4A locus, generated via non-homologous end joining after Cas9 cleavage. The polyclonal format captures diverse editing outcomes, eliminating clonal selection bias and providing a robust model for investigating KDM4A-dependent processes. These cells enable functional analysis without residual gene expression typical of knockdown methods.
The host cell line, HEK293T, is a human embryonic kidney epithelial line derived from HEK293 cells that stably expresses the SV40 large T antigen. This immortalized adherent line supports episomal replication of plasmids containing the SV40 origin, achieving high recombinant protein expression and efficient viral production. Widely used in molecular biology, HEK293T cells are a preferred system for transient gene expression, lentiviral packaging, and functional genomic screens, and their well-characterized signaling networks make them an optimal platform for chromatin and epigenetic research.
KDM4A encodes a histone demethylase that specifically removes di- and trimethyl marks from H3K9 and H3K36, thereby regulating chromatin dynamics and transcription. The enzyme is activated by upstream factors such as androgen receptor (AR), MYC, E2F1, SRC kinase, and DNA damage-activated ATM/ATR kinases. It interacts with coregulators including HDAC1, SIRT1, HP1??, MTA1, and NCoR/SMRT complexes, and cooperates with p53. KDM4A controls downstream targets like CDKN1A/p21, AR-responsive genes, HOXA genes, and CCND1 by demethylating repressive H3K9me3 at promoters and H3K36me3 during elongation. Loss of KDM4A catalytic activity in knockout cells leads to hypermethylation of these histone marks and dysregulated expression of target gene networks.
In the HEK293T background, this knockout model provides a clean system to dissect KDM4A??s epigenetic functions without endogenous enzyme interference. HEK293T cells are readily transfectable and amenable to manipulation of androgen receptor signaling, enabling studies of KDM4A in AR-dependent transcription. The polyclonal population captures diverse null alleles, minimizing clonal bias and facilitating identification of robust phenotypes in DNA damage response, cell cycle regulation, and chromatin-mediated gene silencing. The model is also suitable for investigating KDM4A??s role in cellular senescence and oncogene-induced transcriptional changes.
These polyclonal knockout cells find broad utility in epigenetic and cancer biology investigations. Representative assays include quantitative Western blotting and immunofluorescence to monitor global H3K9me3 and H3K36me3 levels, RT-qPCR and RNA-seq for transcriptome-wide expression profiling, ChIP-qPCR for locus-specific histone modification analysis, and functional studies such as cell proliferation and DNA damage repair assays (e.g., comet assay, ??H2AX foci formation). They are also well-suited for CRISPR-based functional genomics and drug discovery screens. For additional information, please contact Ascent Research.
