HERP CRISPR Activation Plasmid (h): sc-402787-ACT

Human HERPUD1 encodes homocysteine-inducible endoplasmic reticulum protein (HERP), an ER membrane–associated component of the unfolded protein response that helps maintain proteostasis during ER stress. HERP participates in ER-associated degradation (ERAD) by supporting ubiquitin-dependent turnover of misfolded proteins and coordinating crosstalk between stress sensing and proteasomal clearance. Through roles in UPR signaling, redox balance, and quality control, HERPUD1 expression is frequently used as a readout for perturbations that impact protein folding capacity and secretory pathway homeostasis. Dysregulation of ER stress and ERAD pathways is implicated in contexts such as neurodegeneration, metabolic dysfunction, and tumor cell adaptation, making HERPUD1 a useful node for mechanistic studies of stress resilience.

HERP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HERPUD1 expression without altering the underlying DNA sequence.

HERP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HERPUD1 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 HERPUD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HERP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HERPUD1 locus and enabling the study of HERP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HERP pathway restoration in tumor cells with silenced or reduced HERPUD1 expression.

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