The KDM5B Knockout UM-UC-3 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal cell population derived from the human bladder urothelial carcinoma cell line UM-UC-3, in which the KDM5B gene has been disrupted to create a loss-of-function model. This genetically engineered tool employs CRISPR/Cas9-mediated gene editing to target KDM5B, generating a heterogeneous pool of edited cells that collectively ablate KDM5B expression without isolating a single clone. The polyclonal nature preserves the genetic diversity of the knockout population, reducing clonal bias and more accurately reflecting the variable knockout efficiencies encountered in pooled screens or therapeutic contexts. By eliminating KDM5B function, researchers can interrogate the demethylase-dependent epigenetic mechanisms that govern bladder cancer cell proliferation, apoptosis, and invasive behavior.
UM-UC-3 is an established cell line originating from a male patient with transitional cell carcinoma of the bladder, representing a high-grade, muscle-invasive urothelial carcinoma model. These epithelial cells exhibit aggressive growth properties, anchorage-independent growth, and invasive capacity, making them a standard system for dissecting the molecular underpinnings of advanced bladder cancer. The UM-UC-3 parental line carries mutations and expression profiles typical of the luminal-papillary subtype, including alterations in FGFR3 and PI3K pathway components, which intersect with epigenetic modifiers like KDM5B. Performing the knockout in this well-characterized background ensures that the resulting phenotypic changes can be attributed to KDM5B loss within a clinically relevant setting.
KDM5B (JARID1B) is a histone H3 lysine 4 demethylase that catalyzes the removal of di- and trimethyl groups from H3K4, converting active chromatin marks into silent states and thereby repressing gene transcription. This enzyme integrates signals from upstream regulators such as E2F1, MYC, and the PI3K-AKT pathway, while forming repressive complexes with HDAC1/2, MTA2 (a component of the NuRD complex), and SIN3A. Through these interactions, KDM5B directly silences tumor suppressor genes including CDKN2A (p16INK4a), CDKN1A (p21WAF1), and CDH1 (E-cadherin), as well as pro-apoptotic factors like BBC3. KDM5B also modulates stem cell self-renewal and differentiation by controlling HOX gene clusters, thereby linking chromatin dynamics to both oncogenesis and developmental processes.
In the UM-UC-3 bladder cancer model, KDM5B knockout is predicted to relieve transcriptional repression at tumor suppressor loci, leading to increased H3K4me3 occupancy at promoters of CDKN2A and CDKN1A, accompanied by their reactivation. This derepression can impair cell cycle progression, induce apoptosis, and reduce migratory and invasive properties, ultimately diminishing the tumorigenic potential of the cells. The polyclonal knockout population thus serves as a versatile system to study how loss of an epigenetic silencer reshapes the malignant phenotype, offering a platform to validate KDM5B as a therapeutic target in urothelial carcinoma and to explore synthetic lethal interactions with other chromatin modifiers or signaling kinases.
This product is designed for a wide range of applications in cancer epigenetics and bladder cancer research. Users can perform chromatin immunoprecipitation followed by qPCR (ChIP-qPCR) to assess H3K4me3 enrichment at specific target genes, Western blotting to confirm KDM5B depletion and global H3K4me3 changes, and RT-qPCR to measure derepression of CDKN2A and CDKN1A. Functional assays include MTS proliferation assays, Annexin V apoptosis detection, and flow cytometry for cell cycle analysis, as well as Transwell migration and invasion assays to quantify metastatic potential. Additionally, RNA-sequencing can reveal broad transcriptional reprogramming upon KDM5B loss. For further information or technical assistance, please contact Ascent Research.
