MYH9 CRISPR Activation Plasmid (r): sc-437291-ACT

MYH9 encodes non-muscle myosin IIA, an actin-based motor protein that drives cellular contractility, cytokinesis, and dynamic remodeling of the cortical cytoskeleton in rat cells. Through regulation of actomyosin tension, MYH9 influences focal adhesion turnover, cell polarity, and mechanotransduction pathways that coordinate migration and tissue organization. MYH9 function intersects with RhoA/ROCK signaling and integrin-linked cytoskeletal networks that control membrane trafficking, phagocytosis-like processes, and maintenance of epithelial and endothelial barriers. Altered MYH9 activity is linked to defects in platelet production and hemostasis, renal and auditory phenotypes, and broader cytoskeletal dysregulation relevant to studies of inflammation, fibrosis, and cancer cell invasion models.

MYH9 CRISPR Activation Plasmid (r) provides a targeted, non-destructive approach to upregulating endogenous expression without altering the underlying DNA sequence.

MYH9 CRISPR Activation Plasmid (r) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the 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 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MYH9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native locus and enabling the study of MYH9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MYH9 pathway restoration in tumor cells with silenced or reduced expression.

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