GROα CRISPR Activation Plasmid (h): sc-403256-ACT

CXCL1 encodes the chemokine GROα, a secreted ELR+ CXC ligand that binds CXCR2 to promote neutrophil chemotaxis, endothelial activation, and inflammatory cytokine networks. GROα participates in innate immune signaling cascades including NF-κB and MAPK pathways, shaping leukocyte recruitment and tissue remodeling during acute and chronic inflammation. Dysregulated CXCL1/GROα expression has been linked to inflammatory microenvironments and altered stromal–immune interactions observed across multiple disease contexts, making it a useful molecular node for dissecting chemokine-driven cell migration and paracrine signaling. In vitro and in vivo models often leverage CXCL1 to study gradient sensing, myeloid trafficking, and inflammation-associated changes in epithelial and vascular compartments.

GROα CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CXCL1 expression without altering the underlying DNA sequence.

GROα CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CXCL1 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 CXCL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GROα expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CXCL1 locus and enabling the study of GROα-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GROα pathway restoration in tumor cells with silenced or reduced CXCL1 expression.

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