TMEM8C CRISPR Activation Plasmid (h): sc-402487-ACT

TMEM8C, also known as myomaker, encodes a multi-pass transmembrane protein that is essential for myoblast fusion during skeletal muscle development and regeneration. It functions at the plasma membrane to promote fusogenic activity and coordinates with myogenic regulatory networks and membrane remodeling processes required for multinucleated myotube formation. TMEM8C expression is tightly linked to differentiation programs in muscle cells, making it a useful entry point for studying transcriptional control of myogenesis and cell–cell fusion mechanisms. Dysregulation of myoblast fusion and TMEM8C-associated pathways is relevant to experimental models of muscle wasting and inherited myopathies where impaired muscle formation or repair is a key cellular phenotype.

TMEM8C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEM8C expression without altering the underlying DNA sequence.

TMEM8C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM8C 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 TMEM8C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM8C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM8C locus and enabling the study of TMEM8C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM8C pathway restoration in tumor cells with silenced or reduced TMEM8C expression.

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