eIF3α CRISPR Activation Plasmid (h): sc-404354-ACT

EIF3J encodes a subunit of the eukaryotic translation initiation factor 3 (eIF3) complex, a central regulator of cap-dependent translation initiation that coordinates 40S ribosomal subunit recruitment and start codon selection. Through its role in assembling initiation complexes and modulating mRNA-specific translation, EIF3J influences proteostasis, cell growth, and stress-adaptive translational control. Dysregulation of eIF3 subunits has been linked to altered translational programs that support oncogenic signaling, cell cycle progression, and survival pathways, making EIF3J a useful entry point for studying translation-centric mechanisms in disease-relevant contexts. Investigating EIF3J function can inform how selective translation impacts proliferation, apoptosis, and responses to nutrient and ER stress.

eIF3α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF3J expression without altering the underlying DNA sequence.

eIF3α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF3J 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 EIF3J transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF3α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF3J locus and enabling the study of eIF3α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF3α pathway restoration in tumor cells with silenced or reduced EIF3J expression.

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