MYL2 CRISPR Activation Plasmid (h): sc-402798-ACT

MYL2 encodes the ventricular/cardiac regulatory myosin light chain 2, a Ca2+-responsive component of the myosin II complex that modulates actin–myosin cross-bridge cycling and contractile force generation in striated muscle. Through phosphorylation-dependent control of myosin motor activity, MYL2 contributes to sarcomere organization, cardiomyocyte mechanics, and excitation–contraction coupling pathways that coordinate cardiac output. Altered MYL2 expression or function is implicated in inherited and acquired cardiomyopathies, where disrupted sarcomeric regulation can impact myocardial structure and performance. As a marker and regulator of cardiac muscle differentiation and contractility, MYL2 is widely used in studies of cardiac development, disease modeling, and mechanobiology.

MYL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYL2 expression without altering the underlying DNA sequence.

MYL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYL2 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 MYL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MYL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYL2 locus and enabling the study of MYL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MYL2 pathway restoration in tumor cells with silenced or reduced MYL2 expression.

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