hepcidin CRISPR Activation Plasmid (h): sc-403222-ACT

Human HAMP encodes hepcidin, a liver-derived peptide hormone that serves as a central regulator of systemic iron homeostasis. Hepcidin controls dietary iron absorption and macrophage iron recycling by binding the iron exporter ferroportin (SLC40A1), inducing its internalization and degradation, thereby reducing iron efflux to plasma. Its expression is governed by iron-sensing and inflammatory signaling, including BMP/SMAD pathway input from hemojuvelin and HFE/TFR2 complexes, as well as IL-6–STAT3-mediated acute phase responses. Dysregulated hepcidin activity is linked to altered iron distribution and erythropoietic balance in disorders such as hereditary hemochromatosis, anemia of inflammation, and iron-loading anemias, making HAMP a key node for studying iron metabolism and innate immune crosstalk.

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

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

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