MYL3 CRISPR Activation Plasmid (h): sc-403198-ACT

MYL3 encodes myosin light chain 3, a regulatory component of the myosin II motor complex that modulates actin–myosin cross-bridge cycling and contractile force generation in striated muscle. By tuning myosin ATPase activity and filament interactions, MYL3 contributes to sarcomere organization, calcium-sensitive contractility, and myofibrillar mechanics. Altered MYL3 expression or variants have been linked to cardiomyopathic phenotypes and disrupted cardiac muscle function, making it a useful marker for studying sarcomere biology and contractility-associated signaling. Human MYL3 is therefore relevant for investigating muscle development, mechanical stress responses, and pathways governing cardiomyocyte structure and performance.

MYL3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYL3 expression without altering the underlying DNA sequence.

MYL3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYL3 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 MYL3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MYL3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYL3 locus and enabling the study of MYL3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MYL3 pathway restoration in tumor cells with silenced or reduced MYL3 expression.

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