CLIC4 CRISPR Activation Plasmid (h): sc-403188-ACT

ACKR3 (CXCR-7) encodes an atypical chemokine receptor that binds CXCL12/SDF-1 and CXCL11, functioning primarily as a ligand scavenger and signaling modulator rather than a canonical Gαi-coupled chemotactic receptor. By shaping chemokine gradients and receptor cross-talk, CXCR-7 influences CXCR4-dependent migration, survival, and adhesion programs and impacts downstream pathways including MAPK/ERK and β-arrestin–associated signaling. In human tissues, ACKR3 contributes to vascular and immune cell trafficking dynamics, with frequent investigation in contexts such as inflammation, tumor cell invasion, metastasis biology, and angiogenesis. Its expression and activity are also studied in development and tissue remodeling where chemokine availability is rate-limiting for cell positioning.

CLIC4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CLIC4 expression without altering the underlying DNA sequence.

CLIC4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLIC4 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 CLIC4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CLIC4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLIC4 locus and enabling the study of CLIC4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CLIC4 pathway restoration in tumor cells with silenced or reduced CLIC4 expression.

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