CB1/Cannabinoid Receptor 1/CNR1 CRISPR Activation Plasmid (m): sc-419722-ACT

Mouse Cnr1 encodes cannabinoid receptor 1 (CB1/CNR1), a Gi/o-coupled GPCR that responds to endocannabinoids such as anandamide and 2-arachidonoylglycerol to regulate synaptic transmission and neuroendocrine signaling. CB1 activation typically suppresses adenylyl cyclase and cAMP/PKA signaling, modulates MAPK/ERK pathways, and controls ion channel activity to fine-tune neurotransmitter release and neuronal excitability. In the CNS, CNR1 is central to neuromodulatory processes including reward, nociception, appetite, stress responsivity, and learning and memory, with additional roles in peripheral metabolic and immune-related signaling. Altered CNR1 signaling is frequently studied in the context of neuropsychiatric phenotypes, pain-related pathways, and metabolic dysregulation in mouse models.

CB1/Cannabinoid Receptor 1/CNR1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cnr1 expression without altering the underlying DNA sequence.

CB1/Cannabinoid Receptor 1/CNR1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Cnr1 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 Cnr1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CB1/Cannabinoid Receptor 1/CNR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Cnr1 locus and enabling the study of CB1/Cannabinoid Receptor 1/CNR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CB1/Cannabinoid Receptor 1/CNR1 pathway restoration in tumor cells with silenced or reduced Cnr1 expression.

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