Myosin VI CRISPR Activation Plasmid (h): sc-401815-ACT

Human MYO6 encodes myosin VI, an unconventional actin-based motor that uniquely moves toward the minus end of actin filaments to coordinate endocytosis, vesicle trafficking, and membrane remodeling. Myosin VI participates in clathrin-mediated endocytosis, Golgi and endosomal sorting, autophagy-related trafficking, and maintenance of epithelial polarity, and it can influence cytoskeletal dynamics that shape cell migration. Through interactions with adaptor proteins and signaling complexes, MYO6 contributes to pathways governing intracellular transport and actin organization in multiple cell types. Altered MYO6 expression or function has been linked to phenotypes involving sensory and epithelial homeostasis and has been investigated in contexts of tumor biology where trafficking and motility programs are dysregulated.

Myosin VI CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYO6 expression without altering the underlying DNA sequence.

Myosin VI CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYO6 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 MYO6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Myosin VI expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYO6 locus and enabling the study of Myosin VI-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Myosin VI pathway restoration in tumor cells with silenced or reduced MYO6 expression.

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