eIF2Bβ CRISPR Activation Plasmid (h): sc-404686-ACT

EIF2B2 encodes the β subunit of eukaryotic translation initiation factor 2B (eIF2B), the guanine nucleotide exchange factor that regenerates active eIF2–GTP and thereby supports global protein synthesis. eIF2B is a central node in translational control downstream of the integrated stress response, where phosphorylation of eIF2α reduces eIF2B activity to reprogram translation during ER stress, nutrient limitation, and viral or oxidative insults. Through this regulation, EIF2B2 contributes to proteostasis, stress-adaptive gene expression, and cell fate decisions linked to proliferation and survival. Dysfunction in eIF2B complex components is associated with leukodystrophy spectrum disorders and has been studied in the context of stress signaling, neuroglial homeostasis, and translational dysregulation.

eIF2Bβ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF2B2 expression without altering the underlying DNA sequence.

eIF2Bβ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF2B2 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 EIF2B2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF2Bβ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF2B2 locus and enabling the study of eIF2Bβ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF2Bβ pathway restoration in tumor cells with silenced or reduced EIF2B2 expression.

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