The KDM5B Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human clear cell renal cell carcinoma line 786-O, in which the KDM5B gene has been disrupted. This heterogeneous loss-of-function model avoids clonal bias, making it suitable for robust phenotypic and molecular analyses of KDM5B-dependent processes in kidney cancer.
The parental 786-O cell line is an epithelial model established from a primary clear cell renal adenocarcinoma, retaining hallmark features of ccRCC, including VHL-mutant driven HIF1?? accumulation and mesenchymal traits. Its widespread use in renal cancer research provides a well-characterized background for studying epigenetic perturbations that intersect with hypoxia and chromatin remodeling.
KDM5B encodes a lysine-specific demethylase that removes methyl marks from H3K4me2/3, functioning as a transcriptional repressor. It is regulated by HIF1??, p53, retinoic acid receptors, and miR-137, and interacts with RB1, E2F1, EZH2, HDAC1, and the NURF complex. KDM5B directly represses tumor suppressors such as CDKN1A (p21), CDKN2A (p16), and CDH1 (E-cadherin), as well as RB1 and HOX genes, thereby controlling cell cycle progression, apoptosis, and differentiation. This demethylase activity antagonizes the MLL complex and intersects with retinoic acid receptor and RB1/E2F signaling networks.
In 786-O cells, sustained KDM5B activity maintains silencing of these tumor suppressor genes, promoting proliferation and an EMT phenotype. Knockout-mediated disruption relieves transcriptional repression, leading to re-expression of p21, p16, and E-cadherin, which can attenuate cell growth, migration, and invasion. Given the constitutive HIF1?? activation in VHL-mutant 786-O cells, this model also allows investigation of cross-talk between hypoxia and epigenetic silencing.
These polyclonal knockout cells support multiple applications, including ChIP-qPCR for H3K4me3 profiling, RT-qPCR and Western blotting for target gene derepression, and RNA-seq for transcriptome-wide analysis. Genotyping can be confirmed by Sanger sequencing. Functional assays such as MTS proliferation, colony formation, wound healing, and Transwell invasion enable assessment of proliferation and EMT. Apoptosis and protein expression changes can be measured by flow cytometry and immunofluorescence. The cells are also amenable to drug screening for demethylase inhibitors. For further information or to order, contact Ascent Research.
