eIF2C4 CRISPR Activation Plasmid (h): sc-401658-ACT

AGO4, also known as eIF2C4, encodes Argonaute-4, a member of the Argonaute protein family that binds small RNAs to guide sequence-specific regulation of gene expression. Through RNA-induced silencing complex (RISC)-associated processes, AGO4 contributes to post-transcriptional control mechanisms that shape mRNA stability and translation in the cytoplasm. Argonaute-dependent RNA regulation influences pathways governing cell identity, differentiation, and stress responses, and altered small-RNA networks are frequently studied in the context of oncogenic signaling and immune regulation. Dysregulation of Argonaute-mediated gene control has been linked in research settings to aberrant transcriptome remodeling observed across multiple disease-relevant cellular phenotypes.

eIF2C4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AGO4 expression without altering the underlying DNA sequence.

eIF2C4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AGO4 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 AGO4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF2C4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AGO4 locus and enabling the study of eIF2C4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF2C4 pathway restoration in tumor cells with silenced or reduced AGO4 expression.

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