α-Syntrophin CRISPR Activation Plasmid (h): sc-404039-ACT

SNTA1 encodes α-Syntrophin, a modular PDZ-domain scaffold that organizes multiprotein complexes at the inner face of the plasma membrane, particularly within the dystrophin-associated glycoprotein complex in striated muscle and at specialized membrane domains in excitable cells. By coupling ion channels, signaling enzymes, and cytoskeletal elements, α-Syntrophin influences membrane stability, channel localization, and downstream signaling processes that shape cellular excitability and stress responses. SNTA1-dependent scaffolding has been linked to regulation of nitric oxide signaling via nNOS-associated complexes and to the spatial control of membrane receptors and transporters. Dysregulation or variant-associated perturbation of SNTA1 is studied in the context of cardiac electrophysiology, neuromuscular biology, and mechanisms that alter membrane signaling microdomains.

α-Syntrophin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SNTA1 expression without altering the underlying DNA sequence.

α-Syntrophin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SNTA1 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 SNTA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous α-Syntrophin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SNTA1 locus and enabling the study of α-Syntrophin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of α-Syntrophin pathway restoration in tumor cells with silenced or reduced SNTA1 expression.

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