CKR-5 CRISPR Activation Plasmid (h): sc-402548-ACT

CCR5 encodes the human chemokine receptor CKR-5, a seven-transmembrane GPCR predominantly expressed on T cells, macrophages, and dendritic cells where it binds ligands such as CCL3, CCL4, and CCL5 to direct leukocyte chemotaxis. CKR-5 signaling engages heterotrimeric G proteins to activate pathways including PI3K/AKT, PLCβ-driven calcium flux, MAPK/ERK, and cytoskeletal remodeling that coordinate cell migration and inflammatory responses. Through its role in immune cell trafficking and activation, CCR5 is widely studied in inflammation and tissue immune surveillance, and genetic or expression differences have been associated with variation in susceptibility and progression across multiple immune-mediated and infectious disease contexts. In experimental systems, CCR5 expression is often used as a readout for receptor-mediated signaling, chemokine gradient sensing, and modulation of myeloid and lymphoid cell behavior.

CKR-5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCR5 expression without altering the underlying DNA sequence.

CKR-5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCR5 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 CCR5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CKR-5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCR5 locus and enabling the study of CKR-5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CKR-5 pathway restoration in tumor cells with silenced or reduced CCR5 expression.

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