mGluR-3 CRISPR Activation Plasmid (h): sc-404012-ACT

GRM3 encodes the human metabotropic glutamate receptor 3 (mGluR-3), a class C GPCR that modulates synaptic transmission by sensing extracellular glutamate and coupling primarily to Gi/o proteins. Receptor activation inhibits adenylyl cyclase, reduces cAMP signaling, and shapes downstream PKA-dependent responses, while also influencing MAPK/ERK and PI3K-related pathways that affect neuronal excitability and plasticity. mGluR-3 is expressed in neurons and glia, where it contributes to presynaptic control of neurotransmitter release and regulation of neuroinflammatory signaling. Altered GRM3 activity or expression has been investigated in the context of neuropsychiatric and neurodegenerative disease biology, making it a useful target for mechanistic studies of glutamatergic signaling networks.

mGluR-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GRM3 expression without altering the underlying DNA sequence.

mGluR-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GRM3 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the GRM3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous mGluR-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GRM3 locus and enabling the study of mGluR-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of mGluR-3 pathway restoration in tumor cells with silenced or reduced GRM3 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.