eIF2C CRISPR Activation Plasmid (h): sc-409888-ACT

Human AGO1 (eIF2C1) encodes an Argonaute family protein that is a core component of the RNA-induced silencing complex (RISC), coupling small RNAs such as miRNAs and siRNAs to sequence-specific repression of target transcripts. Through small RNA–guided binding, eIF2C participates in post-transcriptional gene regulation, impacting mRNA stability and translation and shaping programs linked to differentiation, stress responses, and maintenance of genome regulatory networks. AGO1-dependent RNA silencing intersects with broader epigenetic and transcriptional control by modulating key signaling nodes and feedback loops that govern cell identity. Dysregulated Argonaute/miRNA pathway activity has been associated with altered gene expression landscapes observed in diverse cancers and neurological and inflammatory conditions, making AGO1 a useful handle for dissecting RNA regulatory mechanisms.

eIF2C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AGO1 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 AGO1 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 AGO1 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 AGO1 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 AGO1 expression.

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