G3BP2 CRISPR Activation Plasmid (h): sc-403630-ACT

Human G3BP2 (G3BP stress granule assembly factor 2) is an RNA-binding protein that acts as a core nucleator of cytoplasmic stress granules, integrating signaling and mRNA metabolism during cellular stress. It participates in translational control, mRNA stabilization/decay, and innate stress responses through interactions that influence ribonucleoprotein complex dynamics and pathways linked to stress-activated kinases. G3BP2 also supports broader post-transcriptional regulation by coordinating RNA-protein assemblies that affect cell survival and adaptation to environmental cues. Dysregulated G3BP2 expression and stress granule biology have been associated with cancer-related phenotypes and neurodegeneration-relevant proteostasis disturbances, making it a useful node for mechanistic studies of RNA granules.

G3BP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous G3BP2 expression without altering the underlying DNA sequence.

G3BP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the G3BP2 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 G3BP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous G3BP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native G3BP2 locus and enabling the study of G3BP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of G3BP2 pathway restoration in tumor cells with silenced or reduced G3BP2 expression.

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