GBP1 CRISPR Activation Plasmid (h): sc-401778-ACT

Human GBP1 (guanylate binding protein 1) is an interferon-inducible large GTPase that functions as an antimicrobial and inflammatory effector downstream of JAK–STAT signaling. GBP1 associates with intracellular membranes and pathogen-containing compartments to regulate cell-autonomous immunity, including inflammasome-associated responses, restriction of microbial replication, and modulation of cytoskeletal and vesicular processes. Through its role in interferon-stimulated gene networks, GBP1 helps shape innate immune transcriptional programs that influence antigen presentation and cytokine-driven signaling. Dysregulated GBP1 expression is frequently used as a marker of heightened interferon activity and has been linked to inflammatory pathobiology and tumor-immune microenvironment states in diverse disease contexts.

GBP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GBP1 expression without altering the underlying DNA sequence.

GBP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GBP1 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 GBP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GBP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GBP1 locus and enabling the study of GBP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GBP1 pathway restoration in tumor cells with silenced or reduced GBP1 expression.

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