eIF3η CRISPR Activation Plasmid (h): sc-400817-ACT

EIF3B encodes eukaryotic translation initiation factor 3 subunit eta (eIF3η), a core component of the eIF3 complex that coordinates assembly of the 43S preinitiation complex and promotes mRNA recruitment to the 40S ribosomal subunit. Through regulation of translation initiation, EIF3B influences proteome remodeling during cell growth, stress responses, and cell cycle progression, and it integrates signaling inputs that shape global and transcript-selective protein synthesis. Dysregulated translation control involving eIF3 subunits has been linked to altered proliferation programs and cellular transformation, making EIF3B a useful node for studying ribosome loading, mRNA selectivity, and proteostasis. Researchers commonly interrogate EIF3B to connect changes in initiation factor function with pathway-level shifts in gene expression outputs.

eIF3η CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF3B expression without altering the underlying DNA sequence.

eIF3η CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF3B 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 EIF3B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF3η expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF3B locus and enabling the study of eIF3η-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF3η pathway restoration in tumor cells with silenced or reduced EIF3B expression.

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