The ID3 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-engineered polyclonal population derived from the 786-O cell line, exhibiting targeted disruption of the ID3 locus. This polyclonal format preserves genetic diversity, avoiding clonal biases and enabling robust interrogation of ID3 function across a heterogeneous cellular background. The cells are provided as an actively growing pool, ready for expansion and immediate use in a range of functional assays, offering a versatile tool for studying gene knockout effects in a cancer cell context.
The 786-O cell line is a human epithelial model originating from clear cell renal cell carcinoma (ccRCC), characterized by inactivating VHL mutations that lead to constitutive HIF stabilization and chronic hypoxic pathway activation. This genetic setting recapitulates key oncogenic features of ccRCC, including dysregulated angiogenesis and metabolic shifts. Consequently, 786-O cells provide a clinically relevant platform for exploring molecular mechanisms in renal cancer and for evaluating potential therapeutics, making them ideal for knockout studies of relevant genes.
ID3 acts as a dominant-negative inhibitor of basic helix-loop-helix (bHLH) transcription factors, principally TCF3 and TCF12, blocking their ability to activate differentiation-associated genes. It occupies a central position in multiple signaling networks, transcriptionally regulated by upstream factors such as TGFB1, BMP4, NOTCH1, WNT3A, HIF1A, EGF, and MYC. ID3 suppresses downstream targets including CDKN1A, CDKN2A, and indirectly upregulates VEGFA, thereby promoting proliferation, survival, and angiogenesis. Through direct interactions with TCF3, TCF12, NEUROD1, MYOD1, and RB1, ID3 sequesters these factors away from promoter regions. Key pathway mediators like SMAD1/5/2/3, NICD/CSL, and MYC integrate signals that converge on ID3 expression, underscoring its role as a hub coordinating growth and differentiation inputs.
In 786-O cells, ID3 overexpression is common due to HIF1A and MYC activation, contributing to a dedifferentiated and aggressive tumor phenotype. CRISPR/Cas9-mediated knockout of ID3 relieves repression of bHLH factors, leading to increased expression of differentiation markers such as CDKN1A, and attenuation of proliferation, migration, and angiogenic capacity. This polyclonal knockout model captures the heterogeneous nature of tumor cell populations, enabling studies on how ID3 loss affects various subpopulations. It serves as a potent system to dissect ID3’s contribution to ccRCC pathogenesis and to test hypotheses regarding differentiation therapy.
These ID3 knockout cells are suited for a broad panel of functional and mechanistic analyses, including proliferation and transwell migration assays, tube formation for angiogenesis assessment, and apoptosis detection by flow cytometry. Transcriptomic profiling via RNA-seq can reveal ID3-dependent gene networks, while co-immunoprecipitation studies may clarify altered protein complexes. They are also valuable for drug screens targeting differentiation or angiogenesis pathways. For further details, custom inquiries, or technical assistance, please reach out to Ascent Research.
