RGS1 CRISPR Activation Plasmid (h): sc-404874-ACT

RGS1 (regulator of G protein signaling 1) is a GTPase-activating protein that accelerates termination of GPCR-driven signaling by promoting Gα subunit inactivation, thereby shaping the magnitude and duration of chemokine and inflammatory cues. In immune cells, RGS1 helps tune migration, activation thresholds, and cytokine-associated responses by modulating pathways downstream of chemokine receptors and other GPCR inputs that converge on second-messenger and MAPK signaling. Altered RGS1 expression has been linked to immune dysregulation and inflammatory disease susceptibility, consistent with its role in controlling leukocyte trafficking and activation programs. As a human gene with strong immunobiology relevance, RGS1 is frequently studied in the context of cell-state transitions, tissue infiltration, and signaling network rewiring.

RGS1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RGS1 expression without altering the underlying DNA sequence.

RGS1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RGS1 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 RGS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RGS1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RGS1 locus and enabling the study of RGS1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RGS1 pathway restoration in tumor cells with silenced or reduced RGS1 expression.

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