utrophin CRISPR Activation Plasmid (h): sc-400522-ACT

Human UTRN encodes utrophin, a dystrophin-related cytoskeletal protein that links the actin network to the dystrophin–glycoprotein complex at the sarcolemma and at specialized membrane domains such as the neuromuscular junction. Utrophin contributes to membrane stability, mechanotransduction, and organization of adhesion-associated signaling by coordinating interactions among extracellular matrix components, transmembrane receptors, and intracellular scaffolds. Its expression is developmentally regulated and often enriched at synaptic regions, supporting studies of muscle fiber architecture and synapse maintenance. Altered utrophin abundance or localization is frequently examined in the context of muscular dystrophy biology and compensatory pathways that influence muscle integrity.

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

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

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