eIF2C CRISPR Activation Plasmid (h): sc-400813-ACT

Human AGO2 (eIF2C2) encodes Argonaute-2, the catalytic core of the RNA-induced silencing complex (RISC) that binds small RNAs to direct sequence-specific repression of target transcripts. AGO2 mediates post-transcriptional gene regulation through miRNA- and siRNA-guided mRNA cleavage, translational repression, and deadenylation, shaping programs involved in cell cycle control, differentiation, innate immune signaling, and stress responses. As a central effector of RNA interference, AGO2 influences transcriptome stability and epigenetic regulation via small-RNA pathways. Dysregulated AGO2 activity or altered miRNA–AGO2 loading has been linked to abnormal gene expression networks observed across multiple disease-relevant contexts, including oncogenic transformation and neurodevelopmental phenotypes, supporting its use in mechanistic studies of RNA silencing.

eIF2C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AGO2 expression without altering the underlying DNA sequence.

eIF2C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AGO2 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 AGO2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF2C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AGO2 locus and enabling the study of eIF2C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF2C pathway restoration in tumor cells with silenced or reduced AGO2 expression.

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