RGS16 CRISPR Activation Plasmid (h): sc-403744-ACT

RGS16 (Regulator of G protein Signaling 16) encodes a GTPase-activating protein that attenuates heterotrimeric Gα subunit signaling to constrain GPCR-driven second messenger pathways, including cAMP and MAPK/ERK outputs. By accelerating GTP hydrolysis on Gαi/o and related G proteins, RGS16 shapes signal duration and amplitude in processes such as chemokine responses, inflammatory signaling, and cell cycle control. Altered RGS16 expression has been reported across multiple disease contexts, reflecting its role in regulating proliferative and immune-associated transcriptional programs. These properties make RGS16 a useful node for dissecting GPCR pathway rewiring and downstream phenotypes in human cell models.

RGS16 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RGS16 expression without altering the underlying DNA sequence.

RGS16 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RGS16 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 RGS16 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RGS16 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RGS16 locus and enabling the study of RGS16-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RGS16 pathway restoration in tumor cells with silenced or reduced RGS16 expression.

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