The ADRM1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the ADRM1 gene. This population consists of a heterogeneous pool of HEK293T cells carrying diverse loss-of-function mutations, avoiding clonal selection artifacts. ADRM1 encodes Rpn13, the 26S proteasome non-ATPase regulatory subunit that functions as a ubiquitin receptor.
HEK293T is a human embryonic kidney cell line that stably expresses the SV40 large T antigen, enhancing transfection efficiency and enabling high-level protein expression and viral production. Its robust growth and compatibility with standard transfection methods make it a versatile platform for studying protein quality control and stress responses. Kidney cells naturally engage active proteasomal pathways, providing a physiologically relevant background for investigating proteasome biology.
Rpn13 captures polyubiquitin chains on substrate proteins and docks them at the 26S proteasome for degradation. It also recruits the deubiquitinating enzyme UCHL5, which edits ubiquitin chains prior to substrate commitment. ADRM1 expression is transcriptionally regulated by NRF1, a factor activated by proteotoxic stress and cytokines. Upon ADRM1 knockout, ubiquitin-proteasome system impairment leads to stabilization of key regulatory proteins, including p53, cyclins, and I??B. Consequently, p53-mediated apoptosis and cell cycle arrest, cyclin-dependent cell cycle progression, and NF-??B signaling via I??B degradation are dysregulated. Rpn13 additionally interacts with adhesion molecules and proteasome subunits, integrating degradation with cellular adhesion and architecture.
In HEK293T cells, ADRM1 knockout provides a tractable system to examine how loss of ubiquitin receptor function alters proteasome substrate turnover and downstream signaling. The SV40 large T antigen partially inactivates p53 and Rb, yet cyclin and I??B stabilization phenotypes remain observable. The polyclonal format facilitates population-level assays such as proteasome activity measurements, drug sensitivity screens, and stress-response profiling without clonal bias, while the high transfection efficiency of HEK293T allows complementary expression of wild-type or mutant ADRM1 for rescue experiments.
These cells are suited for studying the ubiquitin-proteasome system, cancer cell biology, neurodegeneration, and proteasome inhibitor drug validation. Researchers can detect substrate accumulation by western blotting for p53, cyclins, or I??B; quantify proteasome activity fluorometrically; analyze cell cycle by flow cytometry; and evaluate cell viability following proteotoxic stress. Co-immunoprecipitation can identify altered Rpn13-associated complexes. For additional information, contact Ascent Research.
