ETEA CRISPR Activation Plasmid (h): sc-403730-ACT

FAF2 encodes the endoplasmic reticulum–resident protein ETEA (also known as UBXD8), a UBX-domain adaptor that links ubiquitinated substrates to the p97/VCP ATPase to support ER-associated degradation (ERAD) and proteostasis. By coordinating extraction and turnover of misfolded or regulated ER proteins, ETEA contributes to unfolded protein response coupling, lipid droplet organization, and metabolic homeostasis. FAF2/ETEA activity intersects with ubiquitin–proteasome pathways and ER–lipid droplet contact dynamics that influence cellular stress resilience. Altered regulation of ERAD and lipid metabolism has been associated with proteotoxic stress states and metabolic dysfunction, making FAF2 a useful node for mechanistic studies in these contexts.

ETEA CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FAF2 expression without altering the underlying DNA sequence.

ETEA CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FAF2 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 FAF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ETEA expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FAF2 locus and enabling the study of ETEA-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ETEA pathway restoration in tumor cells with silenced or reduced FAF2 expression.

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