The KLLN Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the KLLN tumor suppressor gene in the HEK293T human embryonic kidney cell line. This heterogeneous pool enables loss-of-function studies of KLLN-dependent tumor suppression and DNA damage responses without clonal selection artifacts. The model is particularly relevant for cancer biology research, including investigations into breast, prostate, and colorectal cancers, where KLLN is frequently dysregulated.
The HEK293T host cell line is a hypotriploid epithelial derivative that stably expresses the SV40 large T-antigen, facilitating high-efficiency transient transfection and episomal plasmid replication. Widely employed for recombinant protein production, lentiviral packaging, and functional genomics, HEK293T offers robust growth and consistent performance in vitro. Its well-characterized genetic background, derived from human embryonic kidney, ensures reproducible experimental outcomes across diverse applications.
KLLN functions as a p53-inducible DNA-binding protein that directly interacts with proliferating cell nuclear antigen (PCNA) to halt DNA synthesis. Upon genotoxic stress, TP53 transcriptionally activates KLLN, which then binds PCNA and triggers the mitochondrial apoptotic cascade: BAX activation, cytochrome c release, and subsequent caspase-9 and caspase-3 cleavage, culminating in PARP degradation. This mechanism eliminates cells with unrepaired double-strand breaks, establishing KLLN as a critical tumor suppressor downstream of p53.
In the HEK293T context, KLLN knockout permits dissection of p53-independent activities or reconstitution of the p53-KLLN axis via exogenous expression. Despite SV40 large T-antigen-mediated p53 inhibition, the model supports analysis of PCNA-dependent replication control and apoptotic priming. It facilitates comparative studies of cell cycle distribution, DNA repair kinetics (e.g., ??H2AX foci), and chemosensitivity between wild-type and KLLN-null states, providing insights into p53-pathway targeted therapies.
These cells support diverse assays, including apoptosis detection by Annexin V/PI flow cytometry and cleaved caspase-3/PARP Western blotting, cell cycle profiling with propidium iodide, colony formation assays, and PCNA co-immunoprecipitation. RT-qPCR confirms KLLN transcript reduction, while MTT assays enable viability measurements. The polyclonal nature is ideal for high-throughput drug screens and functional genomics. For further details or to discuss custom requirements, contact Ascent Research.
