CKR-7 CRISPR Activation Plasmid (h): sc-402150-ACT

Human CCR7 encodes the chemokine receptor CKR-7, a G protein–coupled receptor that binds CCL19 and CCL21 to regulate directed migration of T cells, dendritic cells, and other leukocyte subsets. CCR7-driven signaling coordinates chemotaxis, adhesion, and cytoskeletal remodeling through pathways including MAPK/ERK, PI3K–AKT, and Rho family GTPases, supporting lymphoid tissue organization and immune surveillance. Dysregulated CCR7 expression has been associated with altered inflammatory trafficking and with metastatic dissemination patterns in multiple malignancies where chemokine gradients influence tumor cell homing. As a result, CCR7 is widely studied in models of immune cell positioning, antigen presentation dynamics, and microenvironment-driven migration programs.

CKR-7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCR7 expression without altering the underlying DNA sequence.

CKR-7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCR7 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 CCR7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CKR-7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCR7 locus and enabling the study of CKR-7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CKR-7 pathway restoration in tumor cells with silenced or reduced CCR7 expression.

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