S-100A16 CRISPR Activation Plasmid (h): sc-406149-ACT

S100A16 encodes S-100A16, a small EF-hand calcium-binding protein in the S100 family that links intracellular Ca²⁺ dynamics to cell signaling and cytoskeletal organization. By modulating protein–protein interactions in a calcium-dependent manner, S-100A16 has been associated with processes including cell motility, differentiation, and stress-responsive transcriptional programs. Dysregulated S100A16 expression has been reported across multiple disease contexts, including cancers and fibrotic or metabolic phenotypes, where it may influence epithelial–mesenchymal plasticity and microenvironmental remodeling. These features make S-100A16 a useful target for studying calcium-regulated signaling networks and phenotypic transitions in human cells.

S-100A16 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous S100A16 expression without altering the underlying DNA sequence.

S-100A16 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the S100A16 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 S100A16 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous S-100A16 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native S100A16 locus and enabling the study of S-100A16-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of S-100A16 pathway restoration in tumor cells with silenced or reduced S100A16 expression.

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