GRK 6 CRISPR Activation Plasmid (h): sc-402061-ACT

Human GRK6 (G protein-coupled receptor kinase 6) is a serine/threonine kinase that phosphorylates activated GPCRs to promote β-arrestin recruitment, receptor desensitization, and endocytic trafficking. By shaping GPCR signaling dynamics, GRK6 influences downstream second-messenger and kinase cascades including cAMP/PKA and MAPK/ERK, affecting cell migration, inflammatory signaling, and neuronal responses. GRK6 has been studied in the context of chemokine receptor regulation and immune cell trafficking, with disease-relevant links reported across neuroinflammatory processes, cardiovascular signaling dysregulation, and cancer-associated GPCR pathway remodeling. Its endogenous expression level and context-specific activity make GRK6 a useful node for interrogating receptor bias, signaling amplitude, and adaptive feedback in GPCR networks.

GRK 6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GRK6 expression without altering the underlying DNA sequence.

GRK 6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GRK6 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 GRK6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GRK 6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GRK6 locus and enabling the study of GRK 6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GRK 6 pathway restoration in tumor cells with silenced or reduced GRK6 expression.

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