GRINA Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human embryonic kidney HEK293T cell line, engineered to disrupt the GRINA gene. This polyclonal pool offers a heterogeneous loss-of-function model suitable for broad functional genomic studies without single-cell clonal selection. The product provides a ready-to-use cellular system for examining the biological consequences of GRINA ablation in a well-characterized host background.
The parental HEK293T cell line is a widely adopted derivative of HEK293 cells, stably expressing the SV40 large T-antigen. This modification facilitates episomal replication of plasmids containing the SV40 origin and significantly enhances transient protein expression, making HEK293T a workhorse for recombinant protein production, viral packaging, and biochemical assays. Its robust growth characteristics and high transfectability render it a versatile platform for gene-edited cell models.
GRINA (Lifeguard) is an integral membrane protein that functions as a regulatory subunit of N-methyl-D-aspartate (NMDA) glutamate receptors. It physically associates with NMDA receptor subunits GRIN1 and GRIN2A/2B to modulate calcium influx and downstream signaling. GRINA exerts anti-apoptotic effects through dual mechanisms: it attenuates excitotoxic calcium overload, thereby preserving mitochondrial integrity, and it heterodimerizes with Bcl-2 family members Bcl?2 and Bax to inhibit caspase?3 activation. Upstream, GRINA is responsive to glutamate, NMDA, growth factors, and calmodulin/CaMKII-mediated signals, and it influences transcription factors such as CREB. Its signaling network converges on pathways implicated in synaptic plasticity, cell survival, and apoptosis regulation.
In the HEK293T context, GRINA knockout provides a simplified cellular environment to dissect NMDA receptor-dependent calcium homeostasis and apoptotic signaling without the complexity of primary neuronal cultures. Although HEK293T cells do not endogenously express functional NMDA receptors, they can be reconstituted by exogenous co?expression of GRIN subunits, enabling controlled studies of receptor assembly, pharmacology, and GRINA??s modulatory role. The high transfection efficiency of HEK293T cells permits robust overexpression or knockdown of pathway components, facilitating precise interrogation of GRINA interactors such as Bcl?2, Bax, and calmodulin.
This knockout model supports a variety of research applications, including screening for modulators of excitotoxicity, validating drug targets in neurological disorders (schizophrenia, bipolar disorder, Alzheimer??s disease, ischemic brain injury), and investigating apoptosis signaling in cancer. Representative assays include co-immunoprecipitation for protein-protein interactions, calcium imaging with Fluo?4 AM to measure intracellular Ca2+ dynamics, Western blotting and RT?qPCR for expression analysis, Annexin V/PI staining and TUNEL for apoptosis quantification, MTT or LDH release assays for viability, and CREB luciferase reporter assays for transcriptional activity. For further information or to discuss customized experimental strategies, please contact Ascent Research.
