S-100A5 CRISPR Activation Plasmid (h): sc-418401-ACT

S100A5 encodes S-100A5, a Ca2+-binding protein in the S100 family that functions as a dynamic intracellular sensor linking calcium flux to changes in protein interactions and cell state. Like other S100 proteins, S-100A5 can influence signaling and cytoskeletal organization by modulating target proteins in a calcium-dependent manner, integrating cues that affect cellular differentiation and stress responses. Expression of S100A5 is enriched in neural tissues and has been used as a marker of neuronal and glial lineage states, supporting studies of neurodevelopmental programs and activity-dependent transcription. Dysregulated S100 family signaling is frequently investigated in inflammation and cancer biology, making S100A5 a useful node for probing calcium-linked regulatory networks in disease-relevant contexts.

S-100A5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous S100A5 expression without altering the underlying DNA sequence.

S-100A5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the S100A5 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 S100A5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous S-100A5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native S100A5 locus and enabling the study of S-100A5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of S-100A5 pathway restoration in tumor cells with silenced or reduced S100A5 expression.

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