eIF4E2 CRISPR Activation Plasmid (h): sc-403245-ACT

EIF4E2 encodes eIF4E2 (also known as 4EHP), a cap-binding translation initiation factor that modulates mRNA-selective protein synthesis by competing with eIF4E and partnering with repressors to control cap-dependent translation. eIF4E2 contributes to post-transcriptional regulation during stress responses, differentiation, and hypoxia, influencing mRNA stability and translational output in pathways linked to oxygen sensing and cellular adaptation. Through interactions with RNA-binding proteins and microRNA-associated complexes, it shapes gene expression programs that affect cell cycle control, metabolism, and survival. Dysregulated eIF4E2-mediated translational control has been implicated in contexts relevant to cancer biology and other diseases where aberrant protein synthesis and stress signaling are prominent.

eIF4E2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF4E2 expression without altering the underlying DNA sequence.

eIF4E2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF4E2 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 EIF4E2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF4E2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF4E2 locus and enabling the study of eIF4E2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF4E2 pathway restoration in tumor cells with silenced or reduced EIF4E2 expression.

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