Sarcospan CRISPR Activation Plasmid (m): sc-421332-ACT

Mouse Sspn encodes sarcospan, a small tetraspanin-like membrane protein that associates with the dystrophin–glycoprotein complex and related sarcoglycan subcomplexes to support sarcolemma stability and efficient force transmission in striated muscle. Sarcospan contributes to membrane organization, protein trafficking, and signaling events linked to extracellular matrix adhesion and cytoskeletal coupling. Altered SSPN function or expression is studied in the context of muscular dystrophy–related pathways, myofiber fragility, and remodeling responses in skeletal and cardiac muscle. As a modulator of membrane complex assembly, Sspn is relevant for investigations of muscle degeneration, regeneration, and mechanotransduction in mouse models.

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

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

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