Myf-6 CRISPR Activation Plasmid (h): sc-401899-ACT

MYF6 encodes Myf-6 (also known as MRF4), a basic helix–loop–helix transcription factor that functions within the myogenic regulatory factor network controlling skeletal muscle lineage commitment and terminal differentiation. Myf-6 coordinates transcriptional programs linked to myofibrillar assembly, muscle fiber maturation, and maintenance of muscle identity through interactions with E-box regulatory elements and cooperative crosstalk with other myogenic factors. Its activity integrates with pathways governing myogenesis and muscle remodeling, including transcriptional circuitry downstream of developmental cues and epigenetic regulation of muscle-specific gene expression. Dysregulation of myogenic transcriptional control involving MYF6 is relevant to studies of neuromuscular development, muscle degeneration/regeneration dynamics, and tumor contexts showing aberrant myogenic differentiation programs such as rhabdomyosarcoma.

Myf-6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYF6 expression without altering the underlying DNA sequence.

Myf-6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYF6 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 MYF6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Myf-6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYF6 locus and enabling the study of Myf-6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Myf-6 pathway restoration in tumor cells with silenced or reduced MYF6 expression.

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