MIP-3α CRISPR Activation Plasmid (h): sc-417242-ACT

CCL20 encodes the chemokine macrophage inflammatory protein-3α (MIP-3α), a secreted ligand for CCR6 that directs chemotaxis of immature dendritic cells, effector/memory T cells, and B cells to sites of epithelial activation. MIP-3α is induced by NF-κB and MAPK signaling downstream of pattern-recognition receptors and inflammatory cytokines, linking innate immune sensing to leukocyte recruitment and mucosal barrier defense. This chemokine axis shapes lymphoid tissue organization, antigen sampling, and Th17-associated immune polarization within the tissue microenvironment. Dysregulated CCL20/CCR6 signaling has been implicated in chronic inflammatory conditions and tumor-associated inflammation, making it relevant for studying immune cell trafficking and inflammatory circuit remodeling.

MIP-3α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCL20 expression without altering the underlying DNA sequence.

MIP-3α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCL20 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 CCL20 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MIP-3α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCL20 locus and enabling the study of MIP-3α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MIP-3α pathway restoration in tumor cells with silenced or reduced CCL20 expression.

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