α_1A-AR CRISPR Activation Plasmid (m): sc-419021-ACT

Mouse Adra1a encodes the alpha1A-adrenergic receptor (α1A-AR), a G protein–coupled receptor that couples predominantly to Gq/11 to activate phospholipase C, increase IP3/DAG signaling, mobilize intracellular Ca2+, and stimulate PKC-dependent responses. α1A-AR signaling modulates smooth muscle tone, vascular resistance, and sympathetic regulation in cardiovascular and urogenital tissues, and it also influences neuronal excitability and synaptic signaling in the central nervous system. Downstream pathway engagement intersects with MAPK/ERK signaling and transcriptional programs controlling contraction, metabolism, and stress responses. Dysregulated adrenergic receptor activity and altered receptor expression have been implicated in models of hypertension, lower urinary tract dysfunction, and neurobehavioral phenotypes, supporting Adra1a as a target for mechanistic studies of sympathetic signaling networks.

α1A-AR CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Adra1a expression without altering the underlying DNA sequence.

α1A-AR CRISPR Activation Plasmid (m) 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.