GDF-8 CRISPR Activation Plasmid (h): sc-400693-ACT

Human MSTN encodes growth/differentiation factor 8 (GDF-8, myostatin), a secreted TGF-β superfamily ligand that functions as a potent negative regulator of skeletal muscle growth. GDF-8 signals primarily through activin type II receptors and downstream SMAD2/3 phosphorylation to constrain myoblast proliferation and differentiation while influencing protein turnover and metabolic gene programs. This axis intersects with pathways controlling myogenesis, extracellular matrix remodeling, and inflammatory signaling in muscle. Dysregulated MSTN/GDF-8 activity has been linked to altered muscle mass phenotypes and is frequently studied in the context of muscle wasting, sarcopenia, and neuromuscular disease biology.

GDF-8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MSTN expression without altering the underlying DNA sequence.

GDF-8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MSTN 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 MSTN transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GDF-8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MSTN locus and enabling the study of GDF-8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GDF-8 pathway restoration in tumor cells with silenced or reduced MSTN expression.

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