α_1D-AR CRISPR Activation Plasmid (h): sc-402904-ACT

ADRA1D encodes the human α1D-adrenergic receptor (α1D-AR), a G protein-coupled receptor that primarily couples to Gq/11 to activate phospholipase C, increase intracellular Ca2+, and stimulate PKC-dependent signaling. Downstream effects include modulation of MAPK/ERK pathways and transcriptional programs that regulate smooth muscle contractility, vascular tone, and autonomic neurotransmission. α1D-AR activity influences cardiovascular and urogenital physiology and is studied in the context of dysregulated adrenergic signaling associated with hypertension, lower urinary tract dysfunction, and other disorders involving altered smooth muscle responsiveness. ADRA1D expression and signaling dynamics also provide a mechanistic readout for GPCR trafficking, desensitization, and receptor-effector coupling in human cells.

α1D-AR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADRA1D expression without altering the underlying DNA sequence.

α1D-AR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADRA1D 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 ADRA1D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous α1D-AR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADRA1D locus and enabling the study of α1D-AR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of α1D-AR pathway restoration in tumor cells with silenced or reduced ADRA1D expression.

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