Calgranulin A CRISPR Activation Plasmid (h): sc-400610-ACT

S100A8 encodes calgranulin A, a calcium-binding S100 family protein that commonly forms the S100A8/S100A9 heterodimer (calprotectin) in myeloid cells. Calgranulin A participates in innate immune signaling and inflammatory amplification through interactions with pattern-recognition receptors such as TLR4 and RAGE, shaping chemotaxis, leukocyte adhesion, and cytokine production. It is tightly linked to neutrophil and monocyte activation programs and is frequently used as a marker of inflammatory myeloid states. Dysregulated S100A8 expression has been associated with chronic inflammatory disorders, tumor-associated inflammation, and myeloid-driven microenvironment remodeling, making it relevant for pathway-focused mechanistic studies.

Calgranulin A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous S100A8 expression without altering the underlying DNA sequence.

Calgranulin A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the S100A8 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 S100A8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Calgranulin A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native S100A8 locus and enabling the study of Calgranulin A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Calgranulin A pathway restoration in tumor cells with silenced or reduced S100A8 expression.

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