GPSM3 CRISPR Activation Plasmid (m): sc-430655-ACT

Mouse Gpsm3 encodes GPSM3, a GoLoco motif–containing regulator of heterotrimeric G-protein signaling that preferentially modulates Gi/o-class Gα subunits and influences GPCR-driven signal propagation. By shaping G-protein activation status and downstream second-messenger pathways, GPSM3 contributes to chemotactic responses, cytoskeletal dynamics, and context-dependent regulation of inflammatory signaling in myeloid and lymphoid lineages. Genetic and functional studies have linked altered GPSM3 activity with immune-mediated disease susceptibility and variation in inflammatory phenotypes, making it relevant for dissecting leukocyte trafficking and immune homeostasis. In mouse models, perturbing Gpsm3 expression is used to clarify how GPCR signaling thresholds impact innate and adaptive immune responses.

GPSM3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Gpsm3 expression without altering the underlying DNA sequence.

GPSM3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Gpsm3 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 Gpsm3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPSM3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Gpsm3 locus and enabling the study of GPSM3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPSM3 pathway restoration in tumor cells with silenced or reduced Gpsm3 expression.

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