Haptoglobin CRISPR Activation Plasmid (h): sc-401468-ACT

HP encodes haptoglobin, a secreted acute-phase glycoprotein that binds free hemoglobin with high affinity to prevent heme-driven oxidative damage and conserve iron following intravascular hemolysis. The haptoglobin–hemoglobin complex is cleared primarily via CD163-positive macrophages, linking HP to innate immune regulation, inflammatory signaling, and redox homeostasis. As part of the acute-phase response, HP expression is modulated by cytokine-mediated pathways, including IL-6/STAT3 and related transcriptional programs that coordinate systemic inflammation. Altered HP abundance and glycoform patterns have been associated with hemolytic states, inflammatory disorders, and cardiometabolic disease phenotypes, supporting its relevance for mechanistic studies of immunometabolism and oxidative stress.

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

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

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