DYNLT1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population featuring targeted disruption of the DYNLT1 gene. The polyclonal format provides a diverse pool of edited genotypes, enabling comprehensive loss-of-function studies. This model permits investigation of DYNLT1 functions in intracellular transport, mitotic spindle dynamics, and signaling regulation within a widely used human cell line.
The HEK293T host cell line is a human embryonic kidney epithelial derivative, transformed with sheared adenovirus 5 DNA and stably expressing SV40 large T antigen to promote episomal replication of transfected plasmids. These cells are widely employed for protein expression, signaling studies, and high-throughput screening due to their robust growth and high transfectability, providing a versatile background for knockout analyses.
DYNLT1 encodes Tctex-1, a light chain of the cytoplasmic dynein motor complex. It functions in retrograde axonal transport, mitotic spindle organization, and GPCR signaling regulation. The protein interacts with dynein intermediate chain, G-protein ?¦? subunits, Trk receptors, and viral proteins, while also binding and sequestering the pro-apoptotic factor Bim. Upstream regulation includes phosphorylation by PKC and cell cycle-dependent assembly, while downstream it controls cargo trafficking, spindle positioning via NDE1 and pericentrin, and apoptosis through Bim release. Key pathway components include DYNC1H1, DCTN1, PAFAH1B1, and NDE1.
Disruption of DYNLT1 in HEK293T cells compromises dynein retrograde transport, which can alter neurotrophin signaling via Trk receptors and perturb organelle distribution. The consequential release of Bim sensitizes cells to apoptotic triggers, while disrupted interaction with G?¦? subunits modulates GPCR signaling pathways. As a result, these knockout cells serve as a platform to examine dynein-dependent processes in a human epithelial context, with direct implications for cancer cell biology and neurodevelopmental disorders.
Researchers can employ this polyclonal knockout population in diverse assays: live-cell imaging tracks retrograde transport deficits in real time, immunofluorescence microscopy reveals mitotic spindle mispositioning, and co-immunoprecipitation probes dynein complex assembly. Western blotting assesses Bim interaction and phosphorylation status. GPCR signaling reporter assays, flow cytometric apoptosis analysis, and viral infection studies further demonstrate the model??s versatility. It is ideally suited for investigating intracellular trafficking, mitosis, apoptosis regulation, and viral?Chost interactions. For additional technical specifications or inquiries, please contact Ascent Research.
