The AAAS Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T cell line, engineered for the disruption of the AAAS gene. This product provides a heterogeneous pool of cells harboring targeted gene disruptions, enabling the study of AAAS loss-of-function in a human cellular context. The polyclonal format offers a robust model for assessing gene function without the need for single-cell cloning, making it suitable for pooled screening and functional assays.
HEK293T cells are a well-established human embryonic kidney epithelial cell line that stably expresses the SV40 large T antigen. This feature facilitates episomal replication of plasmids containing the SV40 origin, making the line highly permissive for transient transfection and viral production. The parental HEK293T line is widely utilized in biomedical research for protein expression, viral packaging, and functional genomics studies. The derivative polyclonal knockout population retains these characteristics while providing a targeted loss of gene function.
The AAAS gene encodes ALADIN, a scaffold nucleoporin that is an integral component of the NUP107-160 subcomplex within the nuclear pore complex (NPC). ALADIN plays a critical role in NPC integrity and selective nucleocytoplasmic transport, interacting with nucleoporins such as NUP107, NUP160, NUP133, and NUP62. It functions downstream of cell cycle regulators and nuclear envelope assembly factors, mediating the nuclear import of transcription factors including SF-1 and the glucocorticoid receptor via interactions with importin alpha/beta. Disruption of AAAS impairs the nuclear translocation of these factors, thereby affecting gene expression programs essential for cell cycle progression and stress responses.
In the HEK293T host cell background, the AAAS knockout model allows investigation of how ALADIN deficiency impacts NPC assembly and function in a rapidly dividing, transfectable cell line. Given HEK293T’s utility in nuclear transport assays and its epithelial origin, this model is particularly valuable for dissecting the molecular consequences of impaired nucleocytoplasmic shuttling. Researchers can explore effects on downstream targets such as transcription factor import, RNA export, and NPC subcomplex formation, connecting these defects to the pathophysiology of triple A syndrome (achalasia-addisonianism-alacrima).
Typical applications include immunofluorescence localization of NPC components, nuclear import assays using fluorescent reporters, co-immunoprecipitation of nucleoporin subcomplexes, and RT-qPCR for gene expression analysis. The model supports screening for small molecules that restore NPC function and studies of adrenal and neurological cell physiology. The polyclonal population is also amenable to hormone secretion assays and cell viability tests under stress conditions. For further inquiries and technical support, please contact Ascent Research.
