The KDM5D Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population of the human clear cell renal cell carcinoma line 786-O, featuring disruption of the Y-chromosome gene KDM5D (JARID1D). This loss-of-function model eliminates the histone demethylase activity specific for H3K4me2/me3. The heterogeneous edited pool captures diverse genomic disruptions, ensuring robust abrogation of KDM5D function for pooled phenotypic analyses.
The 786-O cell line originates from a primary clear cell renal cell carcinoma in a 58-year-old male. These cells carry a biallelic VHL mutation, causing constitutive HIF stabilization and pathway activation. 786-O is a standard model for VHL-HIF axis dysregulation in kidney cancer, providing a relevant genetic background for studying epigenetic modifier interactions.
KDM5D catalyzes removal of H3K4me2/me3, erasing active chromatin marks to repress transcription. Upstream regulators include the androgen receptor and SRY transcription factor. KDM5D forms co-repressor complexes with HDAC1/2, SIN3A, and retinoblastoma protein (RB1), and interacts with PRC2 components. Its demethylase activity influences downstream targets such as HOX gene clusters and cell cycle regulators, linking epigenetic silencing to cellular proliferation and differentiation.
In the VHL-mutant 786-O context, KDM5D knockout provides a unique tool to dissect the contribution of a Y-chromosome histone demethylase to renal cell carcinoma biology. Loss of KDM5D-mediated H3K4 demethylation may lift transcriptional repression at loci relevant to metabolic adaptation, proliferation, and tumor progression, thereby intersecting the constitutively active HIF pathway. The interplay between chromatin remodeling driven by KDM5D and oncogenic signaling downstream of VHL loss can be systematically explored. Furthermore, given its role in male-specific gene expression, this model enables investigation of sex-biased epigenetic regulation in clear cell kidney cancer.
These polyclonal knockout cells are well suited for a range of molecular and functional assays. ChIP-qPCR can quantify locus-specific H3K4me2/me3 changes upon KDM5D loss, complementing transcriptomic analyses via RNA-seq. Co-immunoprecipitation and mass spectrometry enable mapping of KDM5D interaction networks in the renal cancer proteome. Functional assays??including cell proliferation, migration, and xenograft tumor growth??evaluate impacts on tumorigenic phenotypes. High-throughput drug sensitivity screens can identify compounds that exploit or modulate H3K4 methylation status, aiding epigenetic drug discovery. This model connects Y-chromosome epigenetic regulation to VHL-HIF-driven RCC biology. For further information, please contact Ascent Research.
