Myocardin CRISPR Activation Plasmid (m): sc-431931-ACT

Myocd encodes myocardin, a potent transcriptional coactivator that partners with serum response factor (SRF) to regulate CArG box–dependent gene programs controlling smooth and cardiac muscle differentiation. Myocardin integrates signals from RhoA/actin dynamics and MAPK-associated pathways to coordinate contractile apparatus assembly, cytoskeletal organization, and phenotypic switching of vascular smooth muscle cells. Altered MYOCD activity is linked to dysregulated vascular remodeling and cardiac muscle gene expression, making it a useful node for studying mechanisms relevant to hypertension, atherosclerosis, and cardiomyopathy models. In mouse systems, Myocd perturbation provides a tractable way to interrogate transcriptional control of muscle lineage specification and maturation.

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

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

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