Myosin Va CRISPR Activation Plasmid (h): sc-402108-ACT

MYO5A encodes the actin-based motor protein myosin Va, a processive myosin that couples ATP hydrolysis to long-range transport of membrane-bound organelles and protein complexes along actin filaments. In human cells, myosin Va supports vesicle trafficking, endo/exocytic routing, and polarized delivery of cargo, contributing to cytoskeletal organization and spatial control of signaling at the cell periphery. MYO5A-dependent transport intersects with Rab GTPase-regulated pathways and SNARE-mediated membrane fusion to position secretory granules, receptors, and mRNA/protein assemblies. Disruption or dysregulation of MYO5A has been associated with defects in intracellular transport that can impact neuronal function and pigment-related biology, making it relevant for mechanistic studies of trafficking-linked disease phenotypes.

Myosin Va CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYO5A expression without altering the underlying DNA sequence.

Myosin Va CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYO5A 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 MYO5A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Myosin Va expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYO5A locus and enabling the study of Myosin Va-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Myosin Va pathway restoration in tumor cells with silenced or reduced MYO5A expression.

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