Derlin-1 CRISPR Activation Plasmid (h): sc-404762-ACT

DERL1 encodes Derlin-1, an endoplasmic reticulum (ER) membrane component of the ER-associated degradation (ERAD) machinery that facilitates retrotranslocation of misfolded luminal and membrane proteins for ubiquitin–proteasome turnover. Derlin-1 functions in proteostasis control under basal conditions and during ER stress, interfacing with quality control pathways such as the unfolded protein response and degradation of aberrant glycoproteins. By regulating disposal of misfolded or unassembled proteins, DERL1 influences cellular adaptation to secretory burden, redox imbalance, and proteotoxic stress. Altered DERL1 expression has been reported across multiple disease contexts characterized by chronic ER stress and dysregulated protein homeostasis, supporting its utility as a mechanistic node for studying stress signaling and proteasome-linked clearance.

Derlin-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DERL1 expression without altering the underlying DNA sequence.

Derlin-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DERL1 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 DERL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Derlin-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DERL1 locus and enabling the study of Derlin-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Derlin-1 pathway restoration in tumor cells with silenced or reduced DERL1 expression.

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