GBP2 CRISPR Activation Plasmid (h): sc-402140-ACT

Human GBP2 encodes guanylate-binding protein 2, an interferon-inducible large GTPase that localizes to intracellular membranes and supports cell-intrinsic immunity against diverse pathogens. GBP2 participates in innate immune signaling networks downstream of IFN-γ/STAT1 and contributes to inflammasome-associated processes, antimicrobial restriction, and remodeling of pathogen-containing compartments. By shaping interferon-stimulated gene programs and inflammatory outputs, altered GBP2 expression has been associated with immune dysregulation and inflammatory disease contexts, and it is frequently studied as a marker and mediator of interferon-driven responses in cancer and infection models.

GBP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GBP2 expression without altering the underlying DNA sequence.

GBP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GBP2 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 GBP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GBP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GBP2 locus and enabling the study of GBP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GBP2 pathway restoration in tumor cells with silenced or reduced GBP2 expression.

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