ALB/Albumin CRISPR Activation Plasmid (h): sc-400269-ACT

ALB encodes albumin, the most abundant plasma protein synthesized by hepatocytes and secreted into circulation where it maintains colloid osmotic pressure and serves as a major carrier for fatty acids, bilirubin, hormones, and many small molecules. Albumin participates in systemic nutrient and metabolite transport, influences redox balance through cysteine-based thiol buffering, and contributes to extracellular ligand availability that can shape hepatic metabolic signaling. ALB expression is tightly coupled to hepatocyte differentiation programs and liver-specific transcriptional networks, making it a widely used marker for hepatic identity and function. Altered albumin levels and dysregulated ALB transcription are commonly associated with liver dysfunction and are frequently monitored in studies of hepatic injury, fibrosis, and metabolic disease mechanisms.

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

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

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