GRK 2 CRISPR Activation Plasmid (h): sc-400558-ACT

GRK2 (also known as ADRBK1) encodes G protein-coupled receptor kinase 2, a serine/threonine kinase that phosphorylates activated GPCRs to promote β-arrestin recruitment, receptor desensitization, and trafficking. By shaping signal duration and bias, GRK2 modulates second-messenger pathways including cAMP/PKA, phospholipase C–Ca²⁺ signaling, and MAPK/ERK cascades, with downstream effects on cytoskeletal dynamics and inflammatory signaling. GRK2 also interfaces with non-GPCR substrates and scaffolds, linking receptor inputs to cellular migration, metabolism, and stress responses. Dysregulated GRK2 expression or activity has been associated in the literature with altered cardiovascular signaling, metabolic dysfunction, and cancer-related pathways, supporting its use as a mechanistic node in signaling research.

GRK 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GRK2 expression without altering the underlying DNA sequence.

GRK 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GRK2 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 GRK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GRK 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GRK2 locus and enabling the study of GRK 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GRK 2 pathway restoration in tumor cells with silenced or reduced GRK2 expression.

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