The BTBD10 knockout HEK293T polyclonal cells are a CRISPR/Cas9-edited polyclonal knockout cell population, generated by disrupting the BTBD10 gene in HEK293T cells. This loss-of-function model enables the study of BTBD10, a scaffold protein that couples surface receptor activation to Akt signaling. The heterogeneous pool of edited cells provides a practical system for population-based analyses, avoiding clonal artifacts and allowing efficient screening of BTBD10-dependent phenotypes.
HEK293T is an immortalized human embryonic kidney epithelial cell line stably expressing SV40 large T antigen, which enhances transfection efficiency and episomal plasmid replication. These characteristics have made HEK293T a foundational model for biochemical assays, signal transduction studies, and recombinant protein production. Its non-neuronal origin makes it particularly useful for investigating conserved BTBD10 scaffolding functions without the overlay of synaptic specializations.
BTBD10 is a scaffold protein that facilitates Akt phosphorylation upon stimulation by BDNF/TrkB, AMPA receptors, or GPCR ligands. This activation inhibits apoptosis through phosphorylation of BAD and suppression of FOXO transcription factors, while simultaneously activating mTOR to promote cell growth. BTBD10 directly interacts with the AMPA receptor subunit GluR2, various GPCRs, and the Cul3 ubiquitin ligase complex. Key pathway members include PI3K, Akt, mTOR, BAD, FOXO, GluR2, and PSD-95.
In the HEK293T background, BTBD10 knockout enables dissection of its scaffolding role in receptor-coupled Akt signaling without confounding neuronal synaptic architecture. Researchers can assess receptor-mediated Akt activation, monitor apoptosis via cleaved caspase-3, and examine AMPA receptor trafficking dynamics. The polyclonal knockout format minimizes clonal selection bias, providing a robust platform for comparative signaling studies and dose-response analyses.
These BTBD10 knockout polyclonal cells are suitable for western blotting to quantify Akt phosphorylation kinetics following stimulation with BDNF or GPCR agonists, co-immunoprecipitation to detect altered protein complexes, and immunofluorescence localization of GluR2. Functional apoptosis assays using flow cytometry or caspase activity measurements delineate survival roles, while RT-qPCR can profile FOXO-dependent transcriptional changes. The polyclonal population is ideal for high-content screening, antibody validation, and stable complementation experiments. For technical inquiries, contact Ascent Research.
