MYH11 CRISPR Activation Plasmid (h): sc-400695-ACT

MYH11 encodes smooth muscle myosin heavy chain 11, a core component of the actomyosin contractile apparatus that powers force generation and cytoskeletal remodeling in vascular and visceral smooth muscle cells. Through ATP-dependent interaction with actin filaments, MYH11 supports contractility, cell shape control, and mechanotransduction programs that interface with calcium signaling, RhoA/ROCK-mediated actin dynamics, and focal adhesion turnover. Altered MYH11 expression or structural integrity can disrupt smooth muscle differentiation and contractile function, contributing to vascular wall instability and impaired tissue biomechanics. MYH11 is also implicated in oncogenic gene fusions in myeloid malignancies, highlighting its relevance for studying aberrant transcriptional regulation and lineage-specific phenotypes.

MYH11 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYH11 expression without altering the underlying DNA sequence.

MYH11 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYH11 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 MYH11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MYH11 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYH11 locus and enabling the study of MYH11-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MYH11 pathway restoration in tumor cells with silenced or reduced MYH11 expression.

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