ITI-H2 CRISPR Activation Plasmid (h): sc-402683-ACT

Human ITIH2 encodes inter-alpha-trypsin inhibitor heavy chain 2 (ITI-H2), a liver-enriched secreted glycoprotein that contributes to extracellular matrix stabilization through formation of covalent complexes with hyaluronan via the inter-alpha-inhibitor family. By modulating hyaluronan organization and protease inhibitory networks, ITI-H2 influences tissue remodeling, inflammatory microenvironments, and cell–matrix interactions relevant to wound response and stromal biology. Altered abundance of inter-alpha-inhibitor components, including ITIH2, has been associated with dysregulated extracellular matrix turnover in fibrosis and with tumor-associated matrix remodeling in cancer biology. These properties make ITIH2 a useful target for studying hyaluronan-dependent matrix assembly and inflammation-linked changes in extracellular proteostasis.

ITI-H2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ITIH2 expression without altering the underlying DNA sequence.

ITI-H2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ITIH2 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 ITIH2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ITI-H2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ITIH2 locus and enabling the study of ITI-H2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ITI-H2 pathway restoration in tumor cells with silenced or reduced ITIH2 expression.

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