A1BG CRISPR Activation Plasmid (h): sc-403087-ACT

Alpha-1B-glycoprotein (A1BG) encodes a secreted plasma glycoprotein of the immunoglobulin superfamily that is abundant in circulation and detected across multiple tissues. Although its precise molecular function remains incompletely defined, A1BG is implicated in extracellular protein–protein interactions and modulation of immune and inflammatory processes, with reported associations to oxidative stress and proteostasis-related signaling in the extracellular milieu. Altered A1BG expression has been observed in diverse pathological contexts, including metabolic and inflammatory states and several cancers, supporting its use as a biomarker-linked gene for mechanistic studies. In human cell models, perturbing A1BG provides a route to interrogate pathways governing secreted glycoprotein networks, systemic inflammation, and tumor-associated microenvironmental signaling.

A1BG CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous A1BG expression without altering the underlying DNA sequence.

A1BG CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the A1BG 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 A1BG transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous A1BG expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native A1BG locus and enabling the study of A1BG-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of A1BG pathway restoration in tumor cells with silenced or reduced A1BG expression.

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