Myosin XVIIIA CRISPR Activation Plasmid (h): sc-403216-ACT

Human MYO18A encodes myosin XVIIIA, an unconventional actin-associated motor implicated in organizing the cortical cytoskeleton, regulating cell shape, and coordinating intracellular transport and adhesion-related dynamics. Through interactions with F-actin and scaffolding complexes, MYO18A contributes to processes such as vesicle trafficking, Golgi organization, and migration-associated remodeling that intersect with Rho GTPase signaling and mechanotransduction pathways. Altered MYO18A expression and cytoskeletal dysregulation have been explored in the context of invasive cell behavior and aberrant proliferative signaling, supporting its relevance to disease-associated phenotypes. These features make MYO18A a useful node for studying how actin-based motors couple membrane trafficking to cytoskeletal architecture in human cells.

Myosin XVIIIa CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYO18A expression without altering the underlying DNA sequence.

Myosin XVIIIa CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYO18A 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 MYO18A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Myosin XVIIIa expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYO18A locus and enabling the study of Myosin XVIIIa-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Myosin XVIIIa pathway restoration in tumor cells with silenced or reduced MYO18A expression.

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