MYH7B CRISPR Activation Plasmid (h): sc-400621-ACT

MYH7B encodes a sarcomeric myosin heavy chain family member with roles in actin-based motor activity and the organization of contractile machinery in striated muscle lineages. As part of myosin-dependent force generation, MYH7B is functionally linked to cytoskeletal dynamics, ATP-dependent mechanochemical transduction, and pathways that maintain myofibril integrity and muscle cell differentiation. Dysregulation of myosin heavy chain gene programs is commonly studied in the context of cardiomyocyte and skeletal muscle remodeling, where altered contractile gene expression contributes to pathological phenotypes. MYH7B therefore serves as a useful target for investigating transcriptional control of sarcomere components, muscle-specific gene networks, and stress-responsive remodeling processes in human cell models.

MYH7B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYH7B expression without altering the underlying DNA sequence.

MYH7B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYH7B 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 MYH7B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MYH7B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYH7B locus and enabling the study of MYH7B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MYH7B pathway restoration in tumor cells with silenced or reduced MYH7B expression.

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