Edc3 CRISPR Activation Plasmid (h): sc-402917-ACT

Human EDC3 encodes Edc3, an evolutionarily conserved enhancer of mRNA decapping that links the DCP1–DCP2 decapping complex with additional repressors to promote transcript turnover. Edc3 functions in cytoplasmic processing bodies (P-bodies) and participates in post-transcriptional gene regulation by coupling translational repression to 5′-to-3′ mRNA decay, thereby shaping steady-state RNA abundance and stress-responsive expression programs. Through control of RNA stability, EDC3 influences pathways governing proteostasis, innate stress signaling, and cell-state transitions that depend on rapid remodeling of the transcriptome. Dysregulated mRNA decay and P-body dynamics are broadly implicated in cancer biology and neurodegeneration, making EDC3 a useful node for mechanistic studies of RNA metabolism in disease-relevant contexts.

Edc3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EDC3 expression without altering the underlying DNA sequence.

Edc3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EDC3 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 EDC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Edc3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EDC3 locus and enabling the study of Edc3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Edc3 pathway restoration in tumor cells with silenced or reduced EDC3 expression.

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