The KRT8 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human KRT8 gene in HEK293T cells. This heterogeneous population retains diverse editing events across cells, providing a robust and reproducible loss-of-function model without single-cell cloning biases.
The host HEK293T line is a human embryonic kidney epithelial cell line transformed with adenovirus 5 DNA and stably expressing the SV40 large T antigen. This confers exceptionally high transfection efficiency and supports episomal replication, making it a workhorse for viral packaging and recombinant protein production. Its epithelial origin offers a relevant context for investigating keratin biology.
KRT8 encodes keratin 8, a type II intermediate filament protein that obligately pairs with KRT18 to form heterodimers. These filaments confer mechanical resilience by linking desmosomes and hemidesmosomes through interactions with desmoplakin, plakoglobin, and plectin. Transcription is controlled by p63 and Notch1, with further regulation by AP-1 factors downstream of EGF, TGF-??, PKC, and Akt. Functionally, KRT8 suppresses apoptosis by sequestering the BH3-only protein Bim and modulating death receptor signaling via TNFR2 and Fas. KRT8 loss disrupts keratin networks, releases Bim, and sensitizes cells to apoptotic stimuli while impairing epithelial barrier integrity.
In HEK293T cells, KRT8 knockout decouples keratin function from complex tissue architecture, enabling focused dissection of its anti-apoptotic role. The high transfectability allows efficient rescue experiments, mutant expression, or additional gene targeting, facilitating structure?function studies of the KRT8 interactome. The model is particularly suited for analyzing the Bim/Fas/TNFR2 signaling axis and for screening drugs that exploit keratin-dependent survival mechanisms in epithelial cancers.
Typical applications include immunofluorescence visualization of keratin collapse, Annexin V apoptosis assays, co?immunoprecipitation with KRT18, and transwell migration studies. Researchers employ mechanical stress assays and compound sensitivity profiling to model epithelial fragility disorders and drug?induced toxicity. Western blotting for KRT8 and KRT18 confirms knockout and monitors compensatory changes. For additional information, please contact Ascent Research.
