eIF2α CRISPR Activation Plasmid (h): sc-400199-ACT

EIF2S1 encodes the human eukaryotic translation initiation factor 2 alpha subunit (eIF2α), a central regulator of mRNA translation initiation and proteostasis. Phosphorylation of eIF2α at Ser51 integrates signals from the integrated stress response (ISR) via kinases including PERK, GCN2, PKR, and HRI, reducing global protein synthesis while favoring selective translation programs such as ATF4. Through this mechanism, EIF2S1 links ER stress, nutrient deprivation, viral sensing, and oxidative stress to adaptive transcriptional and metabolic remodeling. Dysregulated eIF2α signaling has been implicated across contexts such as neurodegeneration, cancer cell stress tolerance, and inflammatory pathophysiology, making EIF2S1 modulation useful for pathway dissection.

eIF2α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF2S1 expression without altering the underlying DNA sequence.

eIF2α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF2S1 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 EIF2S1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF2α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF2S1 locus and enabling the study of eIF2α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF2α pathway restoration in tumor cells with silenced or reduced EIF2S1 expression.

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