CBLL1 Knockout HEK293T Polyclonal Cells are a polyclonal population of HEK293T cells engineered with CRISPR/Cas9 to disrupt the CBLL1 gene. This gene-edited product provides a versatile loss-of-function model for investigating CBLL1 (Hakai) biology in a human epithelial context. The polyclonal format offers a heterogeneous knockout pool, suitable for broad functional studies without clonal selection artifacts. CRISPR-mediated gene disruption eliminates CBLL1 expression, enabling researchers to study its role in post-translational regulation and cellular processes.
HEK293T cells are a widely employed human embryonic kidney epithelial line derived from HEK293 cells through stable introduction of SV40 large T antigen, which permits episomal replication of plasmids carrying the SV40 origin of replication. This feature, combined with high transfection efficiency, makes HEK293T a preferred host for protein expression, viral production, and transient or stable genetic manipulation. The epithelial origin of these cells retains key characteristics of cell adhesion and junctional complexes, providing a relevant background for studying CBLL1-mediated regulation of E-cadherin and adherens junctions.
CBLL1 encodes the E3 ubiquitin-protein ligase Hakai, a critical regulator of cell adhesion and epithelial-mesenchymal transition (EMT). Upon phosphorylation by SRC-family kinases, CBLL1 binds and ubiquitinates E-cadherin (CDH1), targeting it for proteasomal degradation. This event disrupts adherens junctions, leading to release of ??-catenin (CTNNB1) from the membrane complex. Stabilized ??-catenin translocates to the nucleus, where it associates with TCF/LEF transcription factors to drive expression of EMT-promoting genes such as SNAI1 and ZEB1. CBLL1 also interacts with proteins including CBL, PTK6, and SRC, and functions downstream of receptor tyrosine kinases such as EGFR and MET. Thus, CBLL1 links extracellular signals to changes in cell adhesion and migratory potential through post-translational control of E-cadherin stability.
CBLL1 disruption in HEK293T cells provides a tractable system for dissecting ubiquitin-dependent E-cadherin regulation in a well-characterized epithelial model. Because HEK293T cells form cadherin-based adherens junctions, loss of CBLL1 is expected to impair E-cadherin ubiquitination and degradation, resulting in stabilized cell-cell contacts and altered ??-catenin signaling. This knockout model facilitates analysis of how SRC-mediated phosphorylation and ubiquitination intersect with cell adhesion dynamics, without confounding effects from single-clone variation. Researchers can employ this polyclonal population to interrogate the interplay between junctional integrity and transcriptional programs that govern EMT.
Typical applications include investigation of E-cadherin turnover, EMT induction, cell migration/invasion, and screening for pharmacological modulators of the CBLL1-E-cadherin axis. Compatible assays range from Western blotting for E-cadherin and ??-catenin levels, immunofluorescence localization of junctional proteins, and migration/invasion assays to E-cadherin ubiquitination analyses and co-immunoprecipitation of the CBLL1-E-cadherin complex. RT-qPCR can monitor transcriptional changes in SNAI1, ZEB1, and other EMT markers, while phospho-SRC signaling can be assessed to explore upstream regulation. These cells are also valuable for unbiased substrate identification and functional genomics studies. For technical support, contact Ascent Research.
