The KRT18 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the human KRT18 gene in the 786-O host cell line. This loss-of-function model is generated via CRISPR/Cas9-mediated gene editing, producing a heterogeneous pool of KRT18-disrupted cells. The polyclonal format minimizes clonal selection artifacts and preserves genetic diversity, making it ideal for robust functional screening and pathway analysis. Researchers can use these cells to interrogate KRT18-dependent processes in a consistent renal carcinoma background.
The host cell line, 786-O, is derived from a primary clear cell adenocarcinoma of the kidney and serves as a widely employed model of human clear cell renal cell carcinoma (ccRCC). These cells carry a homozygous VHL mutation (VHL?/?) and express wild-type PTEN, recapitulating key genetic features of ccRCC. They are extensively used to study VHL-HIF signaling, drug sensitivity, and metastatic dissemination. The 786-O background thus provides a clinically relevant context for evaluating KRT18 knockout phenotypes in renal cancer biology.
Keratin 18 (KRT18) is a type I intermediate filament protein that obligately heterodimerizes with keratin 8 (KRT8) to establish the epithelial cytoskeletal network, anchoring to desmosomes via desmoplakin and plectin. Beyond its structural role, KRT18 modulates apoptosis by binding the death domain adaptor TRADD and impacting caspase-3 activation; its cleavage promotes cytochrome c release and engages Bcl-2 family proteins. KRT18 expression is transcriptionally regulated by p53, SP1, and ETS1, and its activity is influenced by TGF-??, EGF, AKT, and PKC. Downstream, KRT18 affects caspase cascades, BID processing, FADD recruitment, and actin remodeling. Knockout of KRT18 disrupts these signaling nodes, removing a critical apoptotic checkpoint and compromising cytoskeletal integrity.
In the context of 786-O VHL-null cells, KRT18 knockout is particularly valuable for dissecting the crosstalk between intermediate filaments and oncogenic pathways. Loss of KRT18 may impair epithelial barrier function, promote epithelial?Cmesenchymal transition (EMT), and shift apoptosis thresholds, all relevant to ccRCC progression and drug resistance. Because KRT18 interacts directly with AKT and MAPK effectors, this model enables study of cytoskeleton-dependent kinase regulation in a PTEN-intact system, offering insights into therapeutic strategies targeting VHL-HIF-keratin axes.
These knockout cells are suitable for diverse experimental applications, including functional characterization of KRT18 in renal cancer, cytoskeletal dynamics via immunofluorescence, apoptosis assays (e.g., Annexin V/PI and cleaved caspase-3 western blot), and migration/invasion studies. Co-immunoprecipitation can confirm loss of KRT8 interaction, while phospho-AKT and MAPK immunoblotting reveals signaling alterations. Additional uses encompass drug sensitivity screening, RNA-seq transcriptomics, and biomarker discovery. Please contact Ascent Research for further technical information.
