creatine kinase-M CRISPR Activation Plasmid (h): sc-401132-ACT

Human CKM encodes creatine kinase-M, a cytosolic phosphagen kinase that catalyzes the reversible transfer of phosphate between ATP and creatine to generate phosphocreatine, thereby buffering cellular ATP and supporting rapid energy turnover. CKM is highly associated with striated muscle bioenergetics and contributes to metabolic homeostasis during contraction, linking energy demand to glycolytic and oxidative phosphorylation pathways. Altered CKM expression or activity is frequently studied in contexts of muscle injury, myopathies, and metabolic stress, where phosphocreatine dynamics can reflect changes in mitochondrial function and cellular energetics. As a marker of muscle lineage and energy metabolism, CKM is also used to probe differentiation state, sarcomeric organization, and metabolic remodeling in model systems.

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

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

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