α_1A-AR CRISPR Activation Plasmid (h): sc-401726-ACT

ADRA1A encodes the human alpha 1A-adrenergic receptor (α1A-AR), a G protein-coupled receptor that primarily couples to Gq/11 to activate phospholipase C, elevate intracellular Ca2+, and stimulate PKC-dependent signaling. Downstream effects include modulation of smooth muscle contractility, secretion, and neuronal excitability through calcium-dependent pathways and MAPK/ERK signaling. α1A-AR participates in catecholamine-mediated regulation of vascular tone and genitourinary smooth muscle function, and its signaling intersects with broader adrenergic stress-response networks. Altered adrenergic receptor expression or signaling dynamics has been implicated in cardiovascular and neurophysiological phenotypes, supporting research into receptor-dependent transcriptional and second-messenger responses.

α1A-AR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADRA1A expression without altering the underlying DNA sequence.

α1A-AR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADRA1A 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 ADRA1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous α1A-AR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADRA1A locus and enabling the study of α1A-AR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of α1A-AR pathway restoration in tumor cells with silenced or reduced ADRA1A expression.

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