Myoferlin CRISPR Activation Plasmid (h): sc-403110-ACT

Human MYOF encodes myoferlin, a large membrane-associated ferlin family protein that regulates vesicle fusion, endocytosis, and membrane repair through Ca²⁺-dependent lipid binding and coordination of trafficking events at the plasma membrane. Myoferlin supports receptor tyrosine kinase stabilization and recycling, linking it to growth factor signaling dynamics such as EGFR-driven pathways and downstream PI3K/AKT and MAPK cascades. It is also implicated in cytoskeletal remodeling, exocytosis, and cell migration, with functional relevance in myogenesis and angiogenic behavior of endothelial cells. Dysregulated MYOF expression has been associated with altered membrane trafficking states observed in cancer cell invasion and metastasis models, making it a useful node for studying signaling plasticity and tumor microenvironment interactions.

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

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

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