RANTES CRISPR Activation Plasmid (h): sc-401806-ACT

CCL5 encodes the chemokine RANTES, a secreted ligand for CCR1, CCR3, and CCR5 that coordinates leukocyte recruitment, adhesion, and activation during inflammatory responses. RANTES signaling engages GPCR-dependent cascades that intersect with chemotaxis, integrin activation, calcium flux, and downstream MAPK and PI3K pathways, shaping immune cell trafficking within tissues. Dysregulated CCL5 expression is associated with chronic inflammatory microenvironments and altered immune surveillance, making it relevant to studies of autoimmunity, infection-associated immune dynamics, and tumor–immune interactions. In human systems, CCL5/RANTES is commonly investigated in the context of myeloid and T cell communication, cytokine networks, and stromal-immune crosstalk.

RANTES CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCL5 expression without altering the underlying DNA sequence.

RANTES CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCL5 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 CCL5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RANTES expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCL5 locus and enabling the study of RANTES-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RANTES pathway restoration in tumor cells with silenced or reduced CCL5 expression.

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