Calcyclin CRISPR Activation Plasmid (h): sc-404300-ACT

S100A6 encodes calcyclin, a small EF-hand Ca2+-binding protein that functions as a calcium sensor linking dynamic Ca2+ signals to changes in protein interactions and cellular behavior. Calcyclin associates with cytoskeletal and membrane-trafficking factors such as annexins and can modulate actin remodeling, cell-cycle progression, and stress-responsive signaling. It is frequently studied in contexts of proliferation, migration, and differentiation, where altered S100A6 expression has been reported across multiple tumor types and in fibrosis- and inflammation-associated tissue remodeling. These properties make S100A6 a useful node for dissecting calcium-dependent regulatory networks and their impact on cell state transitions.

Calcyclin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous S100A6 expression without altering the underlying DNA sequence.

Calcyclin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the S100A6 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 S100A6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Calcyclin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native S100A6 locus and enabling the study of Calcyclin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Calcyclin pathway restoration in tumor cells with silenced or reduced S100A6 expression.

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