eIF4AIII CRISPR Activation Plasmid (h): sc-402660-ACT

EIF4A3 encodes eIF4AIII, a DEAD-box RNA helicase that serves as a core component of the exon junction complex deposited on spliced mRNAs. eIF4AIII coordinates post-transcriptional gene regulation by coupling pre-mRNA splicing to mRNA export, localization, translation control, and nonsense-mediated mRNA decay, thereby shaping transcriptome fidelity and proteostasis. Through its roles in RNA surveillance and splice-dependent quality control, EIF4A3 influences pathways linked to cell-cycle progression, genome maintenance, and stress responses. Dysregulation of exon junction complex activity and NMD has been associated with aberrant isoform usage and altered expression of oncogenic and neurodevelopmental gene networks, making EIF4A3 a relevant node for mechanistic studies of RNA processing defects.

eIF4AIII CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF4A3 expression without altering the underlying DNA sequence.

eIF4AIII CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF4A3 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 EIF4A3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF4AIII expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF4A3 locus and enabling the study of eIF4AIII-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF4AIII pathway restoration in tumor cells with silenced or reduced EIF4A3 expression.

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