α_2B-AR CRISPR Activation Plasmid (h): sc-403430-ACT

ADRA2B encodes the human α2B-AR, an alpha-2 adrenergic G protein–coupled receptor that preferentially couples to Gi/o to inhibit adenylyl cyclase, reduce intracellular cAMP, and modulate downstream PKA-dependent signaling. Receptor activation also engages β-arrestin–linked processes, receptor internalization and desensitization, and can influence MAPK/ERK pathway activity in a context-dependent manner. α2B-AR contributes to regulation of neurotransmitter release, vascular smooth muscle tone, and sympathetic nervous system responses through catecholamine sensing. Dysregulated adrenergic signaling involving ADRA2B has been associated with phenotypes relevant to cardiovascular and neurobehavioral research, supporting its study in models of stress responsivity, autonomic regulation, and receptor signaling bias.

α2B-AR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADRA2B expression without altering the underlying DNA sequence.

α2B-AR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADRA2B 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 ADRA2B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous α2B-AR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADRA2B locus and enabling the study of α2B-AR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of α2B-AR pathway restoration in tumor cells with silenced or reduced ADRA2B expression.

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