MYH1 CRISPR Activation Plasmid (h): sc-400420-ACT

MYH1 encodes myosin heavy chain 1, a fast-twitch skeletal muscle motor protein that powers actin-based contraction through ATP-dependent cross-bridge cycling within the sarcomere. As a core component of the thick filament, MYH1 contributes to muscle fiber-type specification, force generation, and contractile kinetics, integrating with pathways that regulate myofibrillogenesis, calcium handling, and metabolic adaptation to activity. Variation in myosin heavy chain expression patterns is commonly used to study skeletal muscle differentiation, remodeling, and responses to denervation or exercise-mimetic signaling. Dysregulated sarcomeric gene programs, including shifts in MYH1-associated contractile architecture, are relevant to investigations of myopathies and broader neuromuscular dysfunction mechanisms.

MYH1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYH1 expression without altering the underlying DNA sequence.

MYH1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYH1 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 MYH1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MYH1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYH1 locus and enabling the study of MYH1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MYH1 pathway restoration in tumor cells with silenced or reduced MYH1 expression.

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