beta 3 Adrenergic Receptor/ADRB3/β₃-AR CRISPR Activation Plasmid (h): sc-403301-ACT

ADRB3 encodes the human β3-adrenergic receptor, a G protein–coupled receptor that preferentially engages Gs signaling to stimulate adenylyl cyclase, elevate cAMP, and activate PKA-dependent transcriptional and metabolic programs. In adipocytes and other metabolically active cells, ADRB3 contributes to regulation of lipolysis, thermogenic responses, and energy expenditure through cAMP/PKA and downstream CREB-mediated gene expression, with signaling crosstalk to MAPK pathways depending on cellular context. Altered ADRB3 expression or signaling has been studied in the context of obesity and insulin resistance, as well as broader cardiometabolic phenotypes where adrenergic tone influences cellular metabolism and stress responses. Because β3-AR modulates second-messenger dynamics and receptor desensitization mechanisms, it is frequently used to interrogate GPCR trafficking, ligand bias, and metabolic pathway rewiring.

beta 3 Adrenergic Receptor/ADRB3/β3-AR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADRB3 expression without altering the underlying DNA sequence.

beta 3 Adrenergic Receptor/ADRB3/β3-AR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADRB3 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 ADRB3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous beta 3 Adrenergic Receptor/ADRB3/β3-AR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADRB3 locus and enabling the study of beta 3 Adrenergic Receptor/ADRB3/β3-AR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of beta 3 Adrenergic Receptor/ADRB3/β3-AR pathway restoration in tumor cells with silenced or reduced ADRB3 expression.

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