The HDAC6 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population for loss-of-function studies. This product is a mixed pool of HEK293T cells with Cas9-mediated HDAC6 gene disruption. The polyclonal format offers a robust approach for investigating HDAC6-dependent processes without clonal limitations. This model enables dissection of HDAC6 roles in a widely used human cell background.
HEK293T cells are immortalized human embryonic kidney epithelial cells that stably express the SV40 large T antigen. This cell line is extensively employed for transient protein expression, lentivirus production, and various biochemical assays due to its high transfection efficiency and rapid growth. HEK293T cells exhibit an adherent epithelial morphology and are a standard tool in molecular and cellular biology laboratories. Their robust protein expression machinery and ease of culture make them an ideal host for generating gene knockouts, allowing consistent and reproducible analysis of HDAC6 function in a human cell context.
HDAC6 encodes a cytoplasmic histone deacetylase that primarily deacetylates ??-tubulin and HSP90, modulating microtubule stability, protein folding, and cell motility. It drives aggresome formation and autophagy-mediated clearance of misfolded protein aggregates, linking the ubiquitin-proteasome system to autophagic degradation. HDAC6 is regulated by upstream kinases including EGF/EGFR, Aurora A, GSK3??, p38 MAPK, and ERK1/2, and it influences ??-tubulin acetylation, HSP90 chaperone function, cortactin acetylation, and autophagy flux. Key interactors include p97/VCP, PLAA, ubiquitin, dynein motor complex, tau, Parkin, and LRRK2, which coordinate protein quality control and cell migration. Pathway components HDAC6, ??-tubulin, HSP90, cortactin, p97/VCP, ubiquitinated proteins, dynein, and p62/SQSTM1 form a signaling axis implicated in cancer, neurodegeneration, and inflammation.
In the HEK293T background, disruption of HDAC6 provides a versatile platform for exploring its role in aggresome-autophagy pathways and tubulin-dependent processes. Given that HEK293T cells are frequently utilized for studying protein aggregation and degradation, the HDAC6 knockout model enables precise examination of how deacetylase activity impacts the cellular response to proteotoxic stress. Loss of HDAC6 in these cells typically leads to hyperacetylation of ??-tubulin and HSP90, affecting dynein-mediated transport and chaperone function. This tool is particularly valuable for dissecting signaling networks relevant to cancer cell migration, ciliogenesis, and neurodegenerative disease mechanisms, as the HEK293T system supports both basic and translational research applications.
Researchers can use these cells for western blotting of acetylated ??-tubulin and HSP90, immunofluorescence of aggresomes, and transwell migration/invasion assays. They are suited for autophagy flux assays (LC3-II turnover), co-immunoprecipitation for HDAC6 interactors, tubulin acetylation ELISA, and cell viability under MG132 stress. Applications include studying protein aggregation, cancer cell motility, neurodegeneration, and drug target validation. The polyclonal population provides a convenient system for HDAC6 pathway analysis without cloning. For technical information, contact Ascent Research.
