β1-Syntrophin CRISPR Activation Plasmid (h): sc-405993-ACT

SNTB1 encodes β1-syntrophin, a cytoskeletal adaptor within the dystrophin-associated protein complex that scaffolds signaling proteins at the plasma membrane. Through its PDZ domain, β1-syntrophin organizes microdomains that couple membrane receptors and ion channels to actin-rich structures, shaping processes such as cell adhesion, mechanotransduction, and signal transduction in excitable and non-excitable tissues. SNTB1-dependent complexes influence pathways linked to nitric oxide signaling and channel localization, contributing to membrane stability and regulated cellular responses. Dysregulation of dystrophin-complex components and associated scaffolding networks has been implicated in neuromuscular and cardiomyopathic phenotypes, making SNTB1 a relevant target for investigating disease-associated membrane signaling and cytoskeletal organization.

β1-Syntrophin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SNTB1 expression without altering the underlying DNA sequence.

β1-Syntrophin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SNTB1 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 SNTB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous β1-Syntrophin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SNTB1 locus and enabling the study of β1-Syntrophin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of β1-Syntrophin pathway restoration in tumor cells with silenced or reduced SNTB1 expression.

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