MYH7 CRISPR Activation Plasmid (h): sc-400383-ACT

MYH7 encodes the β-myosin heavy chain, a core sarcomeric motor protein that forms thick filaments in striated muscle and converts ATP hydrolysis into mechanical force. Its regulated interaction with actin and myosin light chains supports contractility, sarcomere assembly, and mechanotransduction programs that couple workload to transcriptional and metabolic adaptation. MYH7 expression and isoform balance are tightly linked to cardiac and slow-twitch skeletal muscle physiology, integrating with calcium-handling, hypertrophic signaling, and cytoskeletal remodeling pathways. Genetic variation or dysregulation of MYH7 is strongly associated with inherited cardiomyopathies and contractile dysfunction phenotypes that are widely modeled in human cell systems.

MYH7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYH7 expression without altering the underlying DNA sequence.

MYH7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYH7 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 MYH7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MYH7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYH7 locus and enabling the study of MYH7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MYH7 pathway restoration in tumor cells with silenced or reduced MYH7 expression.

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