MYH10 CRISPR Activation Plasmid (h): sc-401835-ACT

MYH10 encodes non-muscle myosin IIB, an actin-based motor protein that powers contractile force generation and regulates cytoskeletal organization, cell adhesion, and directional migration. Through its role in actomyosin networks, MYH10 supports cytokinesis, tissue morphogenesis, and mechanotransduction, interfacing with pathways that control Rho GTPase signaling, focal adhesion dynamics, and cortical tension. Altered MYH10 expression or myosin II function has been linked to dysregulated cell motility and developmental processes, making it relevant to studies of epithelial remodeling, neurodevelopment, and fibrosis-associated cellular phenotypes. As a core component of the cytoskeleton, MYH10 is frequently investigated in models probing cell shape changes, barrier function, and force-dependent signaling.

MYH10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYH10 expression without altering the underlying DNA sequence.

MYH10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYH10 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 MYH10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MYH10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYH10 locus and enabling the study of MYH10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MYH10 pathway restoration in tumor cells with silenced or reduced MYH10 expression.

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