eIF4AI CRISPR Activation Plasmid (h): sc-402623-ACT

EIF4A1 encodes the human eIF4AI DEAD-box RNA helicase, a core component of the eIF4F translation initiation machinery that unwinds structured 5′ UTRs to facilitate 43S preinitiation complex scanning and start-codon recognition. By regulating cap-dependent translation efficiency, eIF4AI influences proteome output, cell-cycle progression, and stress-adaptive programs, and it functionally intersects with mTOR signaling and integrated stress responses that remodel mRNA translation. Altered control of eIF4A-dependent translation has been linked to dysregulated growth and survival pathways observed in multiple disease contexts, making EIF4A1 a widely used node for probing translational control mechanisms.

eIF4AI CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF4A1 expression without altering the underlying DNA sequence.

eIF4AI CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF4A1 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 EIF4A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF4AI expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF4A1 locus and enabling the study of eIF4AI-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF4AI pathway restoration in tumor cells with silenced or reduced EIF4A1 expression.

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