The HOMER2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T human embryonic kidney epithelial cell line. This product provides a loss-of-function model for studying the scaffold protein HOMER2, which couples group I metabotropic glutamate receptors to intracellular calcium signaling. The polyclonal format, generated through CRISPR/Cas9-mediated target-gene disruption, yields a heterogeneous knockout cell pool suitable for functional studies without single-cell cloning.
The HEK293T host cell line is immortalized by adenovirus E1A and SV40 large T antigen, conferring high transfectability and robust heterologous protein expression. Derived from human embryonic kidney, these cells retain epithelial characteristics and serve as a standard model for recombinant protein production, viral packaging, and dissection of signaling pathways. HEK293T cells endogenously express key components of calcium and NFAT signaling networks, making them an appropriate host for investigating Homer-dependent mechanisms.
HOMER2 acts as an adaptor that organizes signaling complexes at the interface of G protein-coupled receptors and calcium release machinery. It is activated by GRM1/GRM5 receptor stimulation or T-cell receptor engagement and interacts with inositol 1,4,5-trisphosphate receptors (ITPR1) and TRPC calcium channels to promote calcium mobilization. This process drives calcineurin-mediated dephosphorylation of NFAT transcription factors, including NFATC1, facilitating their nuclear translocation and transcriptional activation. HOMER2 also associates with SHANK1 and modulates ERK1/2 signaling, integrating calcium responses with downstream gene expression.
In the HEK293T context, HOMER2 knockout permits clear discrimination between Homer-dependent and Homer-independent calcium signaling. The line’s high transfectability enables reconstitution studies with wild-type or mutant HOMER2, as well as co-expression of pathway partners such as GRM5, ITPR1, or TRPC1. This model is particularly valuable for dissecting the role of Homer scaffolds in store-operated calcium entry (SOCE) mediated by STIM1 and ORAI1, and for isolating HOMER2-specific contributions to NFAT-driven transcriptional programs.
Applications include intracellular calcium imaging using Fluo-4 to monitor mGluR5 agonist-evoked responses or thapsigargin-induced store depletion, NFAT luciferase reporter assays, co-immunoprecipitation to assess protein interactions, and immunofluorescence for subcellular localization studies. The cells are also suited for cell-based screening of small molecules targeting GRM5?CHOMER2 signaling. These polyclonal knockout cells offer a flexible platform for investigating calcium-dependent pathways in a manipulable human cell system. For further information, please contact Ascent Research.
