The ADAM22 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population designed to disrupt the ADAM22 gene in HEK293T cells. This model provides a robust loss-of-function system for investigating ADAM22??s role in synaptic receptor complex assembly. The polyclonal format offers a diverse pool of gene-edited cells, advantageous for functional studies without clonal bias.
HEK293T cells are an SV40 large T-antigen-immortalized human embryonic kidney epithelial line with high transfection efficiency and capacity for viral packaging. Widely used for protein expression and biochemical reconstitution, these cells serve as an ideal platform for studying synaptic adhesion proteins when engineered to express neuronal components, as they lack endogenous neuron-specific synapse-organizing factors.
ADAM22 is a non-catalytic postsynaptic receptor that binds the secreted neuronal protein LGI1 to form a trans-synaptic scaffold critical for stabilizing AMPA-type glutamate receptors (GluA1, GluA2) at excitatory synapses. This interaction requires additional scaffold and adaptor proteins, including PSD-95 (DLG4) and the transmembrane AMPA receptor regulatory protein TARP ??-2 (stargazin, CACNG2). The LGI1-ADAM22 complex also interacts with the related receptor ADAM23, further stabilizing the postsynaptic density. Disruption of this complex leads to reduced synaptic AMPA receptor content, impaired glutamatergic neurotransmission, and severe seizure phenotypes observed in epileptic encephalopathies.
In the HEK293T background, ADAM22 disruption abrogates LGI1 binding and prevents recruitment of downstream effectors, offering a simplified system to reconstitute the synaptic adhesion complex. Co-expression of LGI1, ADAM23, PSD-95, or stargazin in these knockout cells allows dissection of protein interactions and domain mapping, as well as functional rescue experiments to assess the impact of epilepsy-associated mutations.
Researchers can employ this knockout cell model for co-immunoprecipitation of reconstituted LGI1-ADAM22 complexes, quantitative Western blotting to assess ADAM22 expression, and flow cytometry to measure ligand binding. Immunofluorescence microscopy enables visualization of protein colocalization, while surface biotinylation assays and AMPA receptor trafficking reporter systems provide functional readouts for receptor stabilization. These applications make the ADAM22 Knockout HEK293T Polyclonal Cells a valuable tool for investigating epileptogenic mechanisms, screening small molecules that restore LGI1-ADAM22 binding, and validating synaptic receptor trafficking pathways. For further assistance, please contact Ascent Research.
