The ID3 Knockout UM-UC-3 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the UM-UC-3 bladder cancer cell line, in which the ID3 gene has been disrupted. This pooled format provides a heterogeneous knockout model suitable for functional studies of ID3 without the clonal selection artifacts inherent in single-cell-derived lines. The product enables researchers to interrogate the role of ID3 in urothelial carcinoma biology within a polyclonal context, reflecting varied editing events across the cell population.
UM-UC-3 is a human bladder transitional cell carcinoma cell line originally established from a male patient. It is widely employed as a model for bladder cancer research, exhibiting features of high-grade urothelial carcinoma such as rapid proliferation and invasive capacity. The UM-UC-3 line is particularly relevant for studying tumor cell signaling, drug responses, and mechanisms underlying bladder cancer progression, making it an ideal host for targeted gene disruption.
ID3 encodes a dominant-negative inhibitor of basic helix-loop-helix (bHLH) transcription factors, primarily by forming inactive heterodimers with E-proteins such as E12/E47 (TCF3) and TCF4, thereby preventing bHLH-mediated transcription of target genes including CDKN1A (p21) and cyclin D1, thus promoting cell cycle progression and blocking differentiation. ID3 is transcriptionally regulated by TGF-??, BMP4, and SMAD2/3, and its expression is modulated under hypoxia via HIF1A. The ID3 protein integrates signals from TGF-?? superfamily, hypoxia, and growth factor pathways, functioning as a critical node in proliferation, apoptosis, and differentiation control. In bladder cancer, ID3 dysregulation contributes to unchecked cell growth and tumor maintenance.
Disruption of ID3 in UM-UC-3 cells is anticipated to relieve repression of bHLH factors, increasing p21 expression and potentially inducing cell cycle arrest, apoptosis, or differentiation. This loss-of-function model enables dissection of the ID3-dependent regulatory network in basal/squamous-like bladder cancer contexts, where BMP and TGF-?? signaling are often altered. Comparing polyclonal knockout populations to wild-type UM-UC-3 cells, researchers can evaluate impacts on proliferation, invasive migration, colony formation, and chemosensitivity (e.g., cisplatin). Thus, the model facilitates mechanistic studies linking ID3 to oncogenic pathways in urothelial carcinoma.
Typical research applications include western blotting and RT-qPCR to confirm ID3 ablation and downstream target changes, MTT or BrdU assays to assess proliferation, flow cytometry for cell cycle analysis, and wound healing or Transwell assays to measure migration and invasion. Transcriptome-wide analysis via RNA-seq can identify ID3-dependent gene signatures, while drug sensitivity screens (e.g., cisplatin treatment) reveal therapeutic vulnerabilities. This polyclonal knockout population is particularly suited for pooled functional screens and studies requiring a representative distribution of knockout alleles, avoiding biases introduced by single clones. For additional information about validation and handling, please contact Ascent Research.
