HTRA1 Knockout HCT 116 Polyclonal Cells, a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line. These polyclonal cells harbor a targeted disruption of the HTRA1 gene, providing a loss-of-function model for studying tumor suppression mechanisms. The polyclonal format preserves genetic heterogeneity, reflecting a pooled population of edited cells, and is suitable for experiments where clonal variation is not desired.
HCT 116 is an epithelial colorectal adenocarcinoma cell line with microsatellite instability (MSI-high) and MLH1 deficiency, representing a common molecular subtype of colorectal cancer. This genetic background makes HCT 116 particularly valuable for investigating DNA repair defects, TGF-beta signaling, and Wnt pathway activation. The cell line??s well-characterized mutations in key oncogenic pathways provide a relevant context to assess HTRA1??s tumor-suppressive functions.
HTRA1 encodes a serine protease that functions in protein quality control and extracellular matrix remodeling. It suppresses tumor growth by cleaving extracellular matrix components such as fibronectin and by antagonizing TGF-beta signaling. HTRA1 interacts with TGF-beta receptors and latency-associated peptide to modulate SMAD2 and SMAD3 phosphorylation, thereby inhibiting downstream transcriptional programs. HTRA1 expression is regulated by TGF-beta, BMPs, DNA damage, and oxidative stress. It also influences MAPK and AKT signaling cascades, linking it to multiple proliferative and survival pathways. Downstream, HTRA1 promotes degradation of misfolded proteins and modulates IGFBP bioavailability, affecting cell migration and apoptosis.
In HCT 116 cells, which exhibit TGF-beta responsiveness and canonical Wnt activation, HTRA1 knockout is expected to enhance TGF-beta signaling activity and promote epithelial-to-mesenchymal transition (EMT). Loss of HTRA1 in this MSI-high background may accelerate tumorigenic phenotypes, including increased invasion and resistance to apoptosis. This model is particularly relevant for colorectal cancer research, where HTRA1 expression is frequently downregulated and correlates with poor prognosis. By disrupting HTRA1 in a MLH1-deficient, MSI-high setting, researchers can dissect how protease-dependent tumor suppression intersects with DNA mismatch repair defects and inflammatory signaling.
This polyclonal knockout population is designed for advanced biomedical studies, including TGF-beta signaling reporter assays, cell migration and invasion assays, and apoptosis profiling. Investigators can employ western blotting to assess changes in SMAD2/3 phosphorylation, fibronectin cleavage, and MAPK or AKT activation states. The model supports drug sensitivity screens targeting TGFBR1 or TGFBR2 and functional complementation experiments to rescue protease activity. Additionally, it enables exploration of HTRA1??s role in age-related macular degeneration and osteoarthritis research. For further information, please contact Ascent Research.
