dystrophin CRISPR Activation Plasmid (m): sc-420021-ACT

Dmd encodes dystrophin, a large cytoskeletal protein that links the actin network to the dystrophin–glycoprotein complex at the sarcolemma, supporting membrane stability during muscle contraction. In mouse skeletal and cardiac muscle, dystrophin contributes to mechanotransduction, organization of costameres, and maintenance of calcium homeostasis, with downstream effects on inflammation and fibrotic remodeling pathways when expression is reduced. Disruption or reduced function of dystrophin is associated with progressive myofiber damage and impaired muscle integrity, making Dmd a central gene for studying muscle degeneration phenotypes. Dmd-dependent processes are also relevant to modeling changes in extracellular matrix signaling, stress responses, and myonuclear transcriptional programs in neuromuscular disease contexts.

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

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

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