6CKine CRISPR Activation Plasmid (h): sc-403041-ACT

Human CCL21 encodes the chemokine 6CKine, a homeostatic CCR7 ligand that guides dendritic cells and naïve T cells to secondary lymphoid organs and supports lymphoid tissue organization. By establishing chemotactic gradients, CCL21 regulates leukocyte trafficking, lymphatic endothelial–immune cell crosstalk, and positioning of antigen-presenting cells during immune surveillance. This axis interfaces with GPCR signaling, cytoskeletal remodeling, and adhesion programs that shape cell migration and compartmentalization. Dysregulated CCL21/CCR7 signaling has been associated with altered inflammatory cell recruitment and immune microenvironment remodeling in conditions such as chronic inflammation and tumor-associated lymphangiogenesis, making it a useful target for mechanistic immunology studies.

6CKine CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCL21 expression without altering the underlying DNA sequence.

6CKine CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCL21 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 CCL21 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous 6CKine expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCL21 locus and enabling the study of 6CKine-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of 6CKine pathway restoration in tumor cells with silenced or reduced CCL21 expression.

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