The ATXN1 Knockout HEK293T Polyclonal Cells are a heterogeneous population of HEK293T cells edited by CRISPR/Cas9 to disrupt the ATXN1 gene, generating a polyclonal knockout model for studying ATXN1 function. This product provides a ready-to-use system with loss of ATXN1 expression across the population, enabling studies of transcriptional regulation and disease mechanisms without single-cell cloning.
HEK293T cells are a human embryonic kidney epithelial cell line derived from HEK293 cells, originally transformed with sheared adenovirus 5 DNA. They express SV40 large T antigen, facilitating episomal replication of SV40 ori-containing plasmids and enhancing protein expression and viral production. Their epithelial nature, high transfectability, and rapid growth make them versatile hosts for investigating gene function and signaling pathways.
The ATXN1 gene encodes ataxin-1, a polyglutamine-containing transcriptional repressor that forms complexes with CIC and SMRT/N-CoR2. Its activity is regulated by AKT phosphorylation, cAMP/PKA signaling, sumoylation, and ubiquitination, and it interacts with RBM17, ATXN1L, 14-3-3 proteins, CHIP, Usp7, and PQBP1. This complex represses cerebellar development genes, Notch signaling components, and pro-apoptotic targets; downstream effectors include CIC target genes and Notch effector CBF1, linking ataxin-1 to neurodevelopmental gene programs.
In HEK293T cells, ATXN1 knockout provides a simplified model to study ataxin-1 function without neuronal context. These cells enable dissection of ATXN1-dependent transcriptional repression, protein stability, and interactome dynamics, circumventing clonal variation via the polyclonal population. Although lacking the polyglutamine expansion found in SCA1, they are well suited for evaluating normal ataxin-1 activities and interactions disrupted in disease, including CIC-mediated repression and Notch pathway modulation in a tractable epithelial system.
Applications include reporter assays for CIC activity, RNA-seq for transcriptome profiling, and co-immunoprecipitation of ATXN1 interactors. Western blotting and RT-qPCR confirm knockout, while immunofluorescence reveals subcellular distribution. The cells support screening for modifiers of ataxin-1 function, validation of SCA1 therapeutic targets, proteasome inhibition assays for protein turnover, and cell viability studies for stress responses. For additional information, contact Ascent Research.
