The KRT18 Knockout 769-P Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the 769-P human renal epithelial adenocarcinoma cell line, engineered for targeted disruption of the KRT18 gene (keratin 18). This polyclonal knockout model introduces gene-disrupting edits across a heterogeneous cell pool, mirroring the genetic diversity of tumor populations and minimizing clonal selection bias. The product provides a robust platform for loss-of-function studies of KRT18 in a physiologically relevant epithelial context.
The 769-P host cell line is a clear cell renal cell carcinoma (ccRCC) model isolated from a primary renal adenocarcinoma. It retains key epithelial characteristics and is widely employed in renal cancer research to study tumorigenesis, metastasis, and drug responses. The renal epithelial origin makes it particularly relevant for examining keratin intermediate filaments, as KRT18 and its obligate partner KRT8 are highly expressed and critical for maintaining structural integrity and signaling in these cells.
Keratin 18 (KRT18) is a type I intermediate filament protein that forms heteropolymers with keratin 8 (KRT8), providing mechanical support and participating in cytoprotective signaling. KRT18 interacts with the TNFR1 adaptor TRADD, modulating NF-kB activation and apoptosis upon TNF-alpha stimulation. It also associates with 14-3-3 proteins, linking to PI3K/Akt signaling and cell cycle control. Upstream regulators including AP-1, SP1, EGF, and TGF-beta govern KRT18 expression, while downstream effects involve caspase activation and cell cycle progression. Thus, KRT18 integrates structural and signaling functions critical for epithelial homeostasis.
In the 769-P ccRCC model, KRT18 disruption compromises intermediate filament networks, altering mechanical resilience and signaling pathways that underpin malignant phenotypes. Renal carcinomas often exhibit dysregulated keratin expression, and KRT18 has been implicated in apoptosis resistance and metastatic dissemination. This polyclonal knockout enables dissection of TNFR1-mediated signaling involving TRADD, RIPK1, and NF-kB, and their crosstalk with Akt1 survival signals, without clonal bias. It is well-suited for investigating TNF-alpha-induced apoptosis and epithelial-to-mesenchymal transition (EMT) in renal cancer.
Research applications span renal cell carcinoma epithelial biology, intermediate filament dynamics, and cytokine signaling. The knockout model is ideal for TNF-alpha sensitivity assays, apoptosis profiling via Annexin V flow cytometry, and cell migration and invasion studies to assess metastatic potential. Co-immunoprecipitation experiments probe altered binding with KRT8 and 14-3-3 proteins, while Western blotting and immunofluorescence assess keratin network integrity. RT-qPCR monitors KRT18 and KRT8 mRNA levels. The polyclonal nature of the product supports robust, reproducible experiments by minimizing clonal selection artifacts. For further details, please contact Ascent Research.
