eIF3γ CRISPR Activation Plasmid (h): sc-418339-ACT

EIF3H encodes the eukaryotic translation initiation factor 3 subunit gamma (eIF3γ), a core component of the eIF3 complex that coordinates 40S ribosomal subunit recruitment, mRNA loading, and start-codon recognition during cap-dependent translation initiation. By regulating global and transcript-selective protein synthesis, eIF3γ influences cell growth, stress responses, and proteostasis, intersecting with pathways such as mTOR signaling and integrated stress response programs that tune translational output. Altered eIF3 complex function has been linked to dysregulated translation control observed in proliferative and stress-adaptive cellular states, supporting investigation of EIF3H in disease-relevant models where translational reprogramming contributes to phenotype.

eIF3γ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF3H expression without altering the underlying DNA sequence.

eIF3γ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF3H 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 EIF3H transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF3γ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF3H locus and enabling the study of eIF3γ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF3γ pathway restoration in tumor cells with silenced or reduced EIF3H expression.

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