The CBL Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population of HEK293T cells, designed to disrupt the CBL gene. This format yields a heterogeneous mixture of cells with diverse loss-of-function mutations, enabling robust functional studies in a non-clonal setting. It is particularly useful for pooled assays where population-level responses are critical, such as drug screening and signaling interrogation.
HEK293T cells are a human embryonic kidney epithelial line stably expressing SV40 large T antigen, facilitating episomal plasmid replication. Their adherent growth and epithelial morphology render them ideal for protein overexpression, viral packaging, and biochemical analyses. Widely adopted in signal transduction research, these cells provide a reliable platform for studying gene function in a human epithelial context.
CBL is a RING-type E3 ubiquitin ligase and adaptor that attenuates receptor tyrosine kinase (RTK) signaling. Upon RTK activation, CBL is recruited by upstream kinases such as SRC and SYK to catalyze ubiquitination of activated receptors like EGFR, PDGFR, and MET, marking them for proteasomal degradation. As an adaptor, CBL interacts with GRB2 and SH3KBP1 to coordinate endocytosis and signal termination. In T-cell receptor signaling, CBL functions downstream of ZAP70 and SYK. Through these dual roles, CBL limits MAPK/ERK and PI3K/AKT pathway activity, thereby controlling cell proliferation and survival.
In HEK293T cells, CBL knockout removes negative regulation of RTK cascades, making the model highly responsive to growth factor stimulation. This sensitization allows detailed analysis of EGFR degradation kinetics, sustained phospho-ERK and phospho-AKT signaling, and altered endocytic trafficking. Although non-hematopoietic, HEK293T endogenously expresses relevant RTKs, enabling mechanistic studies in an epithelial background. The polyclonal population offers a genetically diverse resource that avoids clonal artifacts, suitable for examining loss-of-function phenotypes analogous to CBL mutations in myeloid malignancies.
Applications include Western blot-based EGFR degradation assays, phospho-protein profiling, cell proliferation and migration studies, and drug sensitivity testing with kinase or proteasome inhibitors. The knockout cells facilitate validation of CBL as a therapeutic target and support CRISPR-based functional genomics screens. For additional inquiries, please contact Ascent Research.
