The ID3 Knockout KYSE-150 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated to disrupt the ID3 gene in the KYSE-150 human esophageal squamous cell carcinoma cell line. This loss-of-function model enables interrogation of ID3’s role in cancer cell proliferation, differentiation, and signaling. The heterogeneous editing events within the population provide a robust system for functional studies without the limitations of clonal isolation.
KYSE-150 cells originate from a poorly differentiated human esophageal squamous cell carcinoma and retain characteristic epithelial morphology and aggressive growth properties. They exhibit active TGF-beta, BMP, and Wnt signaling pathways, and are commonly used in oncology research. This background offers a clinically relevant setting in which to assess the consequences of ID3 knockout on tumor cell behavior and pathway responsiveness.
ID3 is a dominant-negative helix-loop-helix protein that inhibits transcription by sequestering E-proteins (E12/E47, TCF3), preventing their binding to E-box elements. Its expression is regulated by TGF-beta and BMPs through SMAD2/3 and SMAD1/5/8, respectively, as well as by beta-catenin/TCF and Notch intracellular domain. ID3 represses cell cycle inhibitors p21/CDKN1A and p15/CDKN2B, while promoting cyclin D1 and c-Myc. It interacts with transcriptional coactivators p300/CBP and modulates differentiation factors such as MyoD and NeuroD. This positions ID3 at a central hub coordinating growth and differentiation.
In KYSE-150 cells, ID3 knockout relieves inhibition of bHLH activity, leading to restored transcription of pro-differentiation genes. This is anticipated to reduce proliferation, sensitize cells to apoptotic stimuli, and reverse epithelial-mesenchymal transition markers such as E-cadherin and Snail. Consequently, the knockout model is a valuable resource for dissecting ID3-dependent mechanisms in esophageal squamous cell carcinoma pathogenesis, including its role in maintaining stemness and driving invasion.
The ID3 Knockout KYSE-150 Polyclonal Cells are suitable for a range of applications: investigating TGF-beta/BMP and Notch signaling crosstalk, studying EMT, and assessing drug responses. Typical assays include western blotting for ID3 and downstream effectors, RT-qPCR for EMT genes, MTT or BrdU proliferation assays, Annexin V apoptosis assays, and Transwell migration/invasion assays. Transcriptome analysis via RNA-seq and co-immunoprecipitation of bHLH complexes are also applicable. For additional technical information, please contact Ascent Research.
