The AGTPBP1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the AGTPBP1 gene. This heterogeneous knockout pool provides a loss-of-function model for the cytosolic carboxypeptidase CCP1 (AGTPBP1), supplied as a ready-to-use polyclonal population for immediate experimental analysis of tubulin deglutamylation and associated microtubule dynamics.
The host cell line, HEK293T, is a human embryonic kidney epithelial line stably expressing SV40 large T antigen. Derived from HEK293 cells transformed with adenovirus 5 DNA, it permits episomal replication of SV40 ori-containing plasmids. HEK293T is widely used for transient transfection, protein expression, and viral packaging due to its high transfectability and robust protein production. Its genetic tractability makes it an ideal platform for generating gene-edited cell models.
AGTPBP1 encodes CCP1, a metallocarboxypeptidase that removes C-terminal glutamate residues from ??-tubulin, modulating microtubule stability and motor protein interactions. It functions within a network comprising ??-tubulin, ??-tubulin, microtubule-associated proteins (MAPs), and motor proteins kinesin-1 and dynein. Upstream regulators include neuronal transcription factors and MAPK signaling, while downstream effects mediate microtubule stabilization and axonal transport. Disruption of AGTPBP1 leads to ??-tubulin hyperglutamylation, altering microtubule dynamics and impairing kinesin-1- and dynein-driven intracellular trafficking. This mechanism is central to neurodegeneration, with AGTPBP1 mutations linked to infantile-onset neurodegeneration and cerebellar atrophy (CONDCON).
In the HEK293T context, AGTPBP1 knockout provides a robust model to dissect consequences of impaired tubulin deglutamylation in an epithelial background. Though lacking neuronal processes, these cells retain the core microtubule and trafficking machinery, enabling study of deglutamylation-dependent regulation of microtubule dynamics, organelle transport, and cell division. The polyclonal population captures a spectrum of knockout events, reducing clonal artifacts and providing a representative assessment of phenotypic variance. This model correlates AGTPBP1 loss with altered tubulin glutamylation, microtubule architecture, and motor protein function.
This AGTPBP1 knockout polyclonal cell pool supports diverse applications, including mechanistic studies of microtubule post-translational modifications, neurodegenerative disease modeling, and drug screening. Compatible assays encompass western blotting for tubulin glutamylation, immunofluorescence for microtubule morphology, tubulin polymerization assays, live-cell imaging of intracellular trafficking, and co-immunoprecipitation of tubulin?CMAP interactions. By enabling dissection of AGTPBP1 function, these cells accelerate research in cytoskeletal biology and neuropathology. For further details, please contact Ascent Research.
