ERBB2 Knockout HEK293T Cell Line

The ERBB2 Knockout HEK293T Cell Line is a CRISPR/Cas9-edited human cell line in which the ERBB2 (HER2) gene has been disrupted to ablate target protein expression. This knockout model is derived from the well-characterized HEK293T host cell line (Homo sapiens, embryonic kidney epithelial origin) and provides a defined genetic background for investigating ERBB2-dependent signaling and oncogenic mechanisms. The cell line is supplied as a stable knockout population suitable for functional genomics, drug discovery, and cell signaling studies.

The HEK293T host cells are an immortalized human embryonic kidney epithelial line originally derived by stable expression of the SV40 large T-antigen in HEK293 cells. This modification enhances episomal replication and protein production, making HEK293T a workhorse for transient transfection, recombinant expression, and biochemical analyses. Their robust growth, high transfectability, and well-mapped signaling networks render them an ideal chassis for dissecting receptor tyrosine kinase pathways, including those mediated by the ERBB family.

ERBB2 encodes a 185-kDa receptor tyrosine kinase that lacks a direct ligand but is activated through heterodimerization with ligand-bound ERBB family members such as EGFR, ERBB3, and ERBB4. Ligands including EGF and neuregulins (NRG1) stimulate ERBB receptor phosphorylation, leading to recruitment of adaptor proteins GRB2 and SHC1, which couple the receptor to the RAS?CRAF?CMEK?CERK (MAPK) cascade. Concurrently, ERBB2 engagement activates PI3K?CAKT?CmTOR signaling, fostering cell proliferation, survival, and migration. Additional downstream effectors include STAT3 and MYC, while key transcriptional targets include CCND1, BCL2, and VEGF. Chaperones such as HSP90 and kinases like SRC participate in ERBB2 stabilization and transactivation, while CBL mediates receptor ubiquitination and downregulation. Overexpression or gene amplification of ERBB2 drives constitutive signaling and oncogenic transformation in breast, gastric, ovarian, and lung cancers.

In the HEK293T context, ERBB2 knockout eliminates a central node in the ERBB signaling network, enabling researchers to assess the receptor??s specific contributions to downstream effector activation without interference from endogenous protein. While HEK293T cells do not naturally overexpress ERBB2, they express all core pathway components, making them sensitive to exogenous stimuli or engineered expression. This knockout line thus serves as a clean loss-of-function platform for reconstitution experiments, drug selectivity profiling, and identification of ERBB2-specific signaling partners.

Researchers can employ this cell line in a broad array of applications, including cancer signaling research, drug sensitivity screening with agents such as trastuzumab and lapatinib, and functional validation of ERBB2-targeted therapies. Typical assays include Western blotting and RT-qPCR to confirm knockout, phospho-ERK and phospho-AKT analysis to probe pathway activity, cell proliferation (MTT) and apoptosis (Annexin V) assays, migration (Transwell) assays, and co-immunoprecipitation to study protein interactions. The line is also suitable for functional genomics screens and signal transduction dissection. For further information or technical support, please contact Ascent Research.