Get4 CRISPR Activation Plasmid (h): sc-407595-ACT

Human GET4 encodes Get4, a conserved component of the TRC/GET pathway that supports proteostasis by coordinating post-translational handling and membrane insertion of tail-anchored proteins. Get4 interacts with cochaperone modules and partners such as BAG6/SGTA and TRC40/ASNA1 to triage hydrophobic clients, helping direct them toward endoplasmic reticulum targeting or quality-control degradation. Through its role in tail-anchored protein biogenesis, GET4 influences ER homeostasis, membrane trafficking, and cellular stress responses that are frequently perturbed in cancer and neurodegeneration-associated proteostasis imbalance. Altered GET4 activity can therefore be relevant for studying pathways linked to protein aggregation, ER stress signaling, and ubiquitin-mediated turnover.

Get4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GET4 expression without altering the underlying DNA sequence.

Get4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GET4 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 GET4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Get4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GET4 locus and enabling the study of Get4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Get4 pathway restoration in tumor cells with silenced or reduced GET4 expression.

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