HTRA1 Knockout PaTu 8988t Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the PaTu 8988t human pancreatic ductal adenocarcinoma cell line. This loss-of-function model was generated through CRISPR/Cas9-mediated disruption of the HTRA1 gene, rendering the cell population deficient in HTRA1 serine protease activity. The polyclonal format provides a heterogeneous pool of edited cells, suitable for studying gene function without clonal selection artifacts. HTRA1 is a tumor suppressor frequently downregulated in pancreatic cancer, and its knockout enables investigation of its role in TGF-beta signaling, epithelial-mesenchymal transition, and tumor progression.
PaTu 8988t cells are derived from a human pancreatic adenocarcinoma and serve as a well-established in vitro model of pancreatic cancer. These cells harbor activating KRAS G12V and mutant TP53 mutations, which drive oncogenic signaling and genomic instability. The cell line exhibits typical pancreatic cancer features, including rapid proliferation, metastatic potential, and altered signaling pathway activity. This genetic background makes PaTu 8988t particularly relevant for dissecting tumor suppressor functions and therapeutic resistance mechanisms in pancreatic ductal adenocarcinoma.
HTRA1 encodes a serine protease that preferentially degrades misfolded proteins and cleaves components of the TGF-beta signaling pathway. Mechanistically, HTRA1 suppresses TGF-beta signaling by inactivating TGF-beta ligands and receptors such as TGFBR2, thereby attenuating downstream SMAD2/3 phosphorylation and transcriptional responses. It also influences extracellular matrix remodeling through cleavage of substrates like fibronectin and MMPs. HTRA1 expression is regulated by upstream factors including TGF-beta itself, MAPK signaling, miR-21, and oxidative stress. Its activity modulates interconnected pathways including Wnt, PI3K/AKT, and MAPK, positioning HTRA1 as a critical node in balancing proliferative, migratory, and apoptotic signals.
In PaTu 8988t cells with oncogenic KRAS and TP53 mutations, loss of HTRA1 is expected to exacerbate TGF-beta pathway activation, promoting EMT, migration, and invasion. This knockout model thus recapitulates aspects of aggressive pancreatic cancer where HTRA1 is silenced. Researchers can use these polyclonal knockout cells to dissect how HTRA1 loss cooperates with mutant KRAS and TP53 to drive metastasis and therapy resistance. The model is valuable for validating HTRA1 as a tumor suppressor and exploring its role in modulating the tumor microenvironment through ECM degradation.
These HTRA1 knockout cells are suitable for a wide range of functional genomics and cancer biology studies. Typical applications include examining TGF-beta signaling dynamics via SMAD2/3 phosphorylation assays, assessing cell migration and invasion through Boyden chamber or wound-healing assays, and monitoring proliferation and apoptosis under various conditions. The polyclonal nature supports pooled CRISPR screening and RNA-seq to identify HTRA1-dependent gene expression programs. Co-immunoprecipitation can be employed to investigate HTRA1 interactions with TGF-beta ligands or PDZ domain-containing proteins. Additionally, the model facilitates drug target validation in TGF-beta-driven pathways and EMT research. For further information, please contact Ascent Research.
