Sg II CRISPR Activation Plasmid (h): sc-402906-ACT

SCG2 encodes secretogranin II (Sg II), a member of the granin family that localizes to dense-core secretory granules in neuroendocrine and neuronal cells and supports regulated exocytosis. Proteolytic processing of Sg II generates bioactive peptides such as secretoneurin that modulate chemotaxis, neurite outgrowth, and secretory pathway dynamics, linking SCG2 to vesicle biogenesis, granule acidification, and stimulus-coupled secretion. Through these functions, SCG2 activity intersects with neuroendocrine signaling networks and inflammatory cell trafficking pathways. Altered SCG2 expression and granin peptide profiles have been reported in contexts including neuroendocrine tumor biology and neurological and metabolic disease-associated secretory dysfunction, making it a useful marker and mechanistic node for secretion-focused studies.

Sg II CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SCG2 expression without altering the underlying DNA sequence.

Sg II CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SCG2 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 SCG2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sg II expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SCG2 locus and enabling the study of Sg II-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sg II pathway restoration in tumor cells with silenced or reduced SCG2 expression.

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