Junctophilin-2 CRISPR Activation Plasmid (h): sc-401805-ACT

JPH2 encodes junctophilin-2, a structural membrane-tethering protein that stabilizes junctional membrane complexes between the transverse tubule/plasma membrane and sarcoplasmic reticulum in striated muscle. By maintaining close apposition of these membranes, junctophilin-2 supports efficient excitation–contraction coupling and calcium-induced calcium release through functional coupling of voltage-gated Ca²⁺ channels and ryanodine receptors. JPH2 participates in cardiomyocyte and skeletal muscle Ca²⁺ handling, membrane microdomain organization, and stress-responsive remodeling of junctional architecture. Altered JPH2 expression or function has been linked to dysregulated calcium homeostasis and structural remodeling observed in inherited and acquired cardiomyopathies, including contexts associated with arrhythmogenic phenotypes.

Junctophilin-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous JPH2 expression without altering the underlying DNA sequence.

Junctophilin-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the JPH2 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 JPH2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Junctophilin-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native JPH2 locus and enabling the study of Junctophilin-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Junctophilin-2 pathway restoration in tumor cells with silenced or reduced JPH2 expression.

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