The ID3 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human tongue squamous cell carcinoma line CAL-27. In this product, the gene encoding inhibitor of DNA binding 3 (ID3) has been disrupted using CRISPR/Cas9-mediated gene targeting, generating a heterogeneous pool of cells with loss-of-function mutations. This polyclonal format provides a robust model for studying ID3-dependent cellular processes without the clonal selection bottlenecks, preserving the biological diversity of the knockout phenotype. Researchers can dissect the role of ID3 in cancer cell biology using this well-characterized squamous carcinoma background.
The CAL-27 host cell line is an epithelial cancer cell line established from a human tongue squamous cell carcinoma. This adherent cell line is widely employed in head and neck cancer research due to its reproducible growth characteristics and its relevance to tumor biology. CAL-27 cells retain features of squamous differentiation and harbor genetic alterations typical of head and neck squamous cell carcinoma (HNSCC), making them a suitable platform for investigating oncogenic signaling, invasion, and therapeutic responses. The knockout of ID3 in this context allows direct interrogation of the dominant-negative regulation of basic helix-loop-helix (bHLH) transcription factors within an epithelial tumor microenvironment.
ID3 functions as a dominant-negative inhibitor of bHLH transcription factors, including E2A proteins (E12/E47), HEB, and tissue-specific factors such as MyoD and neurogenin. By sequestering these E proteins in inactive heterodimers, ID3 suppresses the transcription of downstream targets like p21 and cyclin D1, thereby promoting cell cycle progression and inhibiting differentiation. ID3 expression is regulated by multiple upstream signals: Notch signaling via the Notch intracellular domain (NICD) that activates HES1, TGF-beta, and BMP ligands. Additionally, the Hippo pathway converges on these networks, positioning ID3 at a node of cross-talk between proliferation and differentiation cues.
Disruption of ID3 in CAL-27 cells relieves the inhibition of E2A and related bHLH factors, potentially restoring p21 expression and impairing cell cycle entry, while also allowing pro-differentiation programs to proceed. This loss-of-function model is particularly relevant for HNSCC, where overexpression of ID proteins has been associated with tumor aggressiveness, cancer stem cell maintenance, and resistance to conventional therapies. By using these polyclonal knockout cells, scientists can examine how ID3 coordinates the balance between self-renewal and differentiation, and how its removal sensitizes carcinoma cells to apoptotic or anti-proliferative signals.
Typical applications encompass proliferation assays (MTT, BrdU) to assess growth defects, flow cytometry for cell cycle and apoptosis profiling, and western blotting to confirm target protein changes. Migration and invasion assays provide insights into metastatic potential, while RNA-seq and RT-qPCR enable transcriptomic mapping of ID3-regulated networks. The knockout cells are also suitable for drug sensitivity screens, co?culture studies, and in vivo xenograft models. For further information or custom requests, please contact Ascent Research.
The ID3 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human tongue squamous cell carcinoma line CAL-27. In this product, the gene encoding inhibitor of DNA binding 3 (ID3) has been disrupted using CRISPR/Cas9-mediated gene targeting, generating a heterogeneous pool of cells with loss-of-function mutations. This polyclonal format provides a robust model for studying ID3-dependent cellular processes without the clonal selection bottlenecks, preserving the biological diversity of the knockout phenotype. Researchers can dissect the role of ID3 in cancer cell biology using this well-characterized squamous carcinoma background. not done yet.
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