eIF2C3 CRISPR Activation Plasmid (h): sc-401908-ACT

Human AGO3 (eIF2C3) encodes an Argonaute family protein that functions as a core component of the RNA-induced silencing complex, coupling small RNAs to sequence-specific repression of target transcripts. Through interactions with miRNAs and other small RNAs, eIF2C3 contributes to post-transcriptional gene regulation that shapes cell state transitions, stress responses, and network-level control of gene expression programs. Argonaute-dependent pathways intersect with chromatin regulation and innate immune sensing of nucleic acids, influencing pathways relevant to proliferation, differentiation, and tissue homeostasis. Dysregulated small-RNA/Argonaute activity has been linked to altered transcriptome regulation in cancer biology and other complex diseases where miRNA networks are perturbed.

eIF2C3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AGO3 expression without altering the underlying DNA sequence.

eIF2C3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AGO3 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 AGO3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF2C3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AGO3 locus and enabling the study of eIF2C3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF2C3 pathway restoration in tumor cells with silenced or reduced AGO3 expression.

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