The GRIP1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney cell line, featuring targeted disruption of the GRIP1 gene. This product provides a versatile loss-of-function model for investigating the roles of the glutamate receptor interacting protein 1 (GRIP1) scaffold in cellular signaling and protein trafficking pathways. Generated via CRISPR/Cas9-mediated gene disruption, the polyclonal population contains a heterogeneous mix of GRIP1 knockout alleles, enabling robust functional analysis without clonal selection biases. The cells are offered in a ready-to-use format for immediate experimental application in biomedical research.
HEK293T cells serve as an ideal host for this knockout model due to their well-characterized epithelial origin, adherent growth, and stable expression of the SV40 large T antigen. This antigen promotes high-level episomal replication of vectors containing the SV40 origin of replication, yielding exceptional transfection efficiency and protein expression capacity. As a result, HEK293T cells are widely employed for recombinant protein production, lentiviral packaging, and functional genomics studies. Their robust growth characteristics and amenability to standard culture conditions make them a convenient platform for CRISPR-mediated gene editing and downstream biochemical assays.
GRIP1 is a multi-PDZ domain scaffold that anchors AMPA receptors at the postsynaptic density by binding the C-termini of GRIA2 and GRIA3. It forms complexes with EphB receptors, PICK1, Liprin-??, GRASP-1, and GIPC1 to coordinate receptor trafficking and synaptic cell adhesion. Upstream regulators such as EphB receptors, PKC, CaMKII, PKA, and NMDA receptor activation modulate GRIP1 function. Downstream, GRIP1 couples AMPA receptors to KIF5 kinesin motors and influences surface expression of GRIA1 and GRIA2, thereby regulating receptor endocytosis and recycling. Through these interactions, GRIP1 is essential for synaptic plasticity and glutamatergic signaling.
Although HEK293T cells lack the full set of neuronal structures, they provide a powerful reductionist system for dissecting GRIP1??s biochemical functions. Disruption of GRIP1 in these cells enables the study of its scaffolding role independent of synaptic environment, facilitating investigation of protein-protein interactions, post-translational modifications, and signaling pathway integration. The knockout background allows researchers to reconstitute specific interactions by expressing wild-type or mutant GRIP1 variants alongside AMPA receptor subunits, EphB2, or other binding partners. This model is particularly valuable for examining how GRIP1 regulates surface trafficking of AMPA receptors using surface biotinylation assays, or for assessing the impact of GRIP1 loss on EphB receptor-mediated signaling cascades.
Typical research applications include screening for pharmacological modulators of glutamatergic signaling, investigating GRIP1 interaction networks via co-immunoprecipitation, and analyzing downstream gene expression by RT-qPCR. The cells are suitable for immunofluorescence localization studies, western blotting, and surface biotinylation assays to track AMPA receptor trafficking. This knockout model supports research on synaptic dysfunction associated with Fraser syndrome, autism spectrum disorders, schizophrenia, and intellectual disability, providing a versatile platform for functional genomic studies. For additional technical information or to place an order, please contact Ascent Research.
