The HTRA1 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population generated from the CAL-27 human tongue squamous cell carcinoma line, featuring targeted disruption of the HTRA1 gene. This product provides a heterogeneous pool of edited cells, without clonal isolation, and serves as a versatile loss-of-function model for investigating the tumor-suppressive functions of HTRA1 in oral cancer biology.
CAL-27 is a well-established human epithelial cell line derived from a tongue squamous cell carcinoma in a male donor. These cells are tumorigenic and widely used as a model for oral squamous cell carcinoma, exhibiting key features of epithelial carcinogenesis such as unregulated proliferation and invasive potential. The parental line??s consistent growth characteristics and tractable genetics make it a robust host for CRISPR-mediated gene disruption studies.
HTRA1 encodes a serine protease that acts as a tumor suppressor through multiple mechanisms, including the cleavage of misfolded proteins, modulation of TGF-?? signaling, and induction of apoptosis. Transcription of HTRA1 is activated by p53 and E2F1 and is responsive to TGF-?? stimulation; epigenetic silencing via promoter methylation is a common event in cancers. The HTRA1 protein assembles as a homotrimer and proteolytically processes substrates such as fibronectin, decorin, and TGF-??1, while also promoting caspase-3-mediated cell death. In the TGF-?? pathway, HTRA1 interacts functionally with TGFBR1 and regulates the phosphorylation of SMAD2/3 and their complex formation with SMAD4, thereby influencing downstream MAPK8 (JNK) and p38 MAPK signaling. Additionally, HTRA1 associates with tubulin, which may link its activity to cytoskeletal dynamics and cell motility.
Knockout of HTRA1 in CAL-27 oral cancer cells is predicted to relieve tumor-suppressive constraints, resulting in enhanced cell proliferation, migration, and resistance to apoptosis. This model permits direct examination of how loss of HTRA1-mediated proteolysis of TGF-??1 and ECM components alters signaling through TGFBR1-SMAD2/3-SMAD4 and MAPK cascades, potentially shifting the cellular phenotype toward greater malignancy. It also allows investigation of the interplay between HTRA1 loss and other oncogenic pathways, such as Wnt signaling, in a clinically relevant background.
Researchers can utilize this polyclonal knockout population in a variety of functional assays, including Western blotting for pathway markers (e.g., phospho-SMAD2, fibronectin, caspase-3), RT-qPCR for gene expression analysis, and TGF-?? signaling reporter assays to quantify pathway activity. Phenotypic characterization can be performed through proliferation, migration/invasion, and apoptosis assays, while co-immunoprecipitation studies can probe HTRA1??s interactions with tubulin or other partners. These cells are also suitable for drug screening efforts targeting the TGF-??, MAPK, or Wnt pathways, and for functional genomics screens aimed at identifying synthetic lethal interactions. For technical inquiries or to obtain a quotation, please contact Ascent Research.
