G3BP1 CRISPR Activation Plasmid (h): sc-400745-ACT

Human G3BP1 (G3BP stress granule assembly factor 1) is an RNA-binding protein that nucleates stress granule formation and coordinates mRNA triage during cellular stress. It integrates signaling from translation control and innate immune pathways by regulating mRNA stability, translation initiation, and ribonucleoprotein remodeling, with downstream effects on apoptosis, antiviral responses, and cell-cycle programs. G3BP1 participates in stress-adaptive processes linked to eIF2α-mediated translational arrest, MAPK signaling, and RNA metabolism, and its dysregulation has been associated with altered stress tolerance and aberrant proliferation in multiple disease contexts. Because G3BP1 couples RNA processing to stress-responsive signaling, it is widely studied in models of tumor biology, neurodegeneration, and host–pathogen interactions.

G3BP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous G3BP1 expression without altering the underlying DNA sequence.

G3BP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the G3BP1 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 G3BP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous G3BP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native G3BP1 locus and enabling the study of G3BP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of G3BP1 pathway restoration in tumor cells with silenced or reduced G3BP1 expression.

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