S-100 α chain CRISPR Activation Plasmid (h): sc-402065-ACT

S100A1 encodes the S-100 α chain, a calcium-binding EF-hand protein that functions as a signaling mediator linking intracellular Ca2+ dynamics to protein–protein interaction networks. In human cells, S100A1 participates in regulation of cytoskeletal organization, cellular motility, and stress-responsive signaling, influencing processes such as myogenesis, mitochondrial function, and excitation–contraction coupling through calcium-dependent pathways. Altered S100A1 expression has been reported across multiple pathological contexts, including cardiovascular dysfunction, neuromuscular remodeling, and cancers, where it can affect proliferation, invasion, and differentiation programs. As a molecular readout of calcium-dependent signaling states, S100A1 is commonly studied in pathway mapping and mechanistic models of tissue-specific homeostasis and disease-associated remodeling.

S-100 α chain CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous S100A1 expression without altering the underlying DNA sequence.

S-100 α chain CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the S100A1 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 S100A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous S-100 α chain expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native S100A1 locus and enabling the study of S-100 α chain-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of S-100 α chain pathway restoration in tumor cells with silenced or reduced S100A1 expression.

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