ATXN2L Knockout HEK293T Polyclonal Cells offer a CRISPR/Cas9-edited polyclonal cell population for disruption of the ATXN2L gene in HEK293T cells. This loss-of-function model avoids clonal variability, providing a robust genetic background for investigating ATXN2L-dependent post-transcriptional regulation. The polyclonal format ensures broad representation of editing events, supporting population-level functional studies in RNA biology and neurodegeneration.
The HEK293T host cell line is derived from human embryonic kidney epithelial cells and stably expresses the SV40 large T antigen. This feature enhances episomal plasmid replication and transfection efficiency, rendering the cells ideal for recombinant protein production, lentiviral packaging, and a wide range of functional assays. Their well-characterized physiology and ease of manipulation make HEK293T a standard platform for dissecting gene function in a controlled in vitro setting.
ATXN2L encodes an RNA-binding scaffold protein that localizes to cytoplasmic stress granules, where it interacts with PABPC1, ATXN2, eIF4E, TIA1, and G3BP1. It is activated by cellular stressors such as oxidative stress and heat shock, downstream of mTOR signaling, to modulate mRNA translation and stability. By influencing translation initiation factors and regulated mRNA targets, ATXN2L plays a central role in RNA metabolism and proteostasis. Knockout of ATXN2L disrupts stress granule dynamics and alters post-transcriptional control, making it a pivotal factor in the mRNA surveillance pathway.
In the HEK293T context, ATXN2L knockout enables clear phenotypic analysis of stress granule biology due to the cells’ robust transfection capability and compatibility with live-cell imaging. The SV40 large T antigen does not hinder stress granule assembly, permitting reproducible studies with agents like arsenite. This model is particularly valuable for rescue experiments and ectopic expression of tagged granule components, facilitating mechanistic dissection of RNA?Cprotein interactions. Although epithelial, HEK293T cells recapitulate conserved RNA regulatory mechanisms relevant to neurodegenerative disorders.
Applications include modeling spinocerebellar ataxia type 2 and amyotrophic lateral sclerosis, employing assays such as western blotting for ATXN2L and stress markers, immunofluorescence for G3BP1 and TIA1, and RT-qPCR for target transcript expression. Co-immunoprecipitation and RNA immunoprecipitation further probe altered protein and RNA interactions. Arsenite-induced stress assays quantify functional defects in stress granule formation. These tools enable comprehensive investigation of ATXN2L’s role in cellular stress responses and translation control. For additional information or custom solutions, please contact Ascent Research.
