MyoD CRISPR Activation Plasmid (h): sc-400092-ACT

MYOD1 encodes the human MyoD basic helix–loop–helix transcription factor that functions as a master regulator of skeletal myogenesis. MyoD binds E-box motifs and cooperates with MEF2 family members and chromatin remodelers to activate muscle lineage programs, coordinate cell-cycle exit, and promote differentiation through transcriptional control of myogenic structural and metabolic genes. Its activity intersects with signaling inputs including Wnt/β-catenin, Notch, TGF-β/SMAD, and MAPK pathways that tune myoblast commitment and maturation. Dysregulated MYOD1 expression or function is used as a marker of altered myogenic state and has been studied in contexts such as impaired regeneration, muscle wasting biology, and MYOD1-associated transcriptional programs in rhabdomyosarcoma models.

MyoD CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYOD1 expression without altering the underlying DNA sequence.

MyoD CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYOD1 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 MYOD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MyoD expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYOD1 locus and enabling the study of MyoD-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MyoD pathway restoration in tumor cells with silenced or reduced MYOD1 expression.

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