STUB1 Knockout HEK293T Cell Line

The STUB1 Knockout HEK293T Cell Line is a CRISPR/Cas9-edited knockout cell line featuring targeted disruption of the STUB1 gene in the HEK293T human embryonic kidney host cell line. This product offers a stable loss-of-function model to investigate STUB1-dependent ubiquitylation and protein quality control mechanisms.

The HEK293T host is an adenovirus 5-transformed epithelial cell line that constitutively expresses the SV40 large T antigen, conferring high transfectability and episomal plasmid replication. Widely used for recombinant protein production and viral packaging, HEK293T provides a robust and scalable platform for gene-edited cell lines.

STUB1 encodes a co-chaperone and E3 ubiquitin ligase that bridges chaperone machinery and the ubiquitin-proteasome system. It directly binds Hsp70/Hsc70 (HSPA8, HSPA1A) and Hsp90 to recognize misfolded clients, and with E2 enzymes UBE2D1/2 and UBE2N, catalyzes substrate ubiquitination for proteasomal degradation. Activity is regulated by BAG-family co-chaperones (BAG1, BAG2, BAG3) and induced by HSF1-mediated transcription under heat shock, oxidative stress, or unfolded protein response. Targets include tau, p53, ErbB2, and CFTR-??F508, linking STUB1 to neurodegeneration, cancer, cardiac diseases, and aging-related proteotoxicity.

In HEK293T cells, STUB1 knockout disrupts the proteostasis network, leading to altered degradation of client proteins. The model is particularly suited to examine chaperone-assisted quality control and to study how loss of E3 ligase function affects protein accumulation and aggregation-prone substrates, without complications of tissue-specific factors.

This cell line enables ubiquitination assays, Western blot-based degradation monitoring, co-immunoprecipitation of Hsp70/Hsp90 complexes, and proteasome inhibition (MG132) experiments to assess proteasome-dependent turnover. Cycloheximide chase analysis can measure protein half-lives, and CETSA provides target engagement profiling. The knockout line supports investigation of how STUB1-mediated ubiquitylation intersects with other degradation pathways. Applications include mechanistic studies of protein quality control, drug target validation in cancer and neurodegeneration, and high-throughput screening for STUB1 activity modulators. For further details, please contact Ascent Research.