myoglobin CRISPR Activation Plasmid (m): sc-421575-ACT

Mouse Mb encodes myoglobin, a heme-containing oxygen-binding protein that buffers and facilitates intracellular O₂ diffusion in striated muscle, supporting oxidative phosphorylation during fluctuating workload. By modulating local oxygen availability and nitric oxide/redox chemistry, myoglobin influences mitochondrial respiration, reactive oxygen species handling, and metabolic adaptation to hypoxia and exercise. Altered myoglobin abundance or function has been investigated in contexts of muscle injury, myopathies, and ischemia-associated stress where oxygen transport and mitochondrial performance are limiting factors. In biomedical research, Mb serves as a tractable node for studying muscle energetics, oxygen-sensing programs, and oxidative stress pathways in vivo and in cultured myocytes.

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

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

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