Edc4 CRISPR Activation Plasmid (h): sc-403803-ACT

Human EDC4 encodes Edc4, a conserved scaffold of cytoplasmic processing bodies (P-bodies) that coordinates mRNA decapping and 5′–3′ decay by organizing core factors such as DCP1/DCP2 and associated exonucleases. Through its role in post-transcriptional gene regulation, Edc4 influences mRNA turnover, translational repression, and stress-responsive ribonucleoprotein granule dynamics. Perturbation of P-body and decapping pathways can remodel transcript stability programs that shape proliferation, differentiation, and inflammatory signaling. Dysregulated RNA metabolism and decapping complex activity have been linked to oncogenic and neurobiological phenotypes, supporting the use of EDC4 as a node for studying disease-relevant RNA homeostasis.

Edc4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EDC4 expression without altering the underlying DNA sequence.

Edc4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EDC4 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 EDC4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Edc4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EDC4 locus and enabling the study of Edc4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Edc4 pathway restoration in tumor cells with silenced or reduced EDC4 expression.

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