Moesin CRISPR Activation Plasmid (h): sc-401065-ACT

Human MSN encodes moesin, a member of the ERM (ezrin–radixin–moesin) family that links cortical F-actin to transmembrane proteins to organize membrane structure and mechanics. Moesin regulates actin cytoskeleton remodeling, cell shape, adhesion, and migration through phosphorylation-dependent activation and participation in Rho GTPase- and PI(4,5)P2-associated signaling. By coordinating microvilli formation, immune cell polarization, and endothelial barrier dynamics, moesin influences processes such as cell trafficking and tissue organization. Dysregulated ERM activity and cytoskeletal signaling are frequently investigated in the context of altered invasiveness, metastatic potential, and aberrant immune cell behavior in disease models.

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

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

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