Pdcd-4 CRISPR Activation Plasmid (h): sc-401743-ACT

PDCD4 encodes Pdcd-4, a tumor suppressor–like protein that modulates translation by interacting with initiation factors and RNA helicases, thereby influencing mRNA-specific protein synthesis and cellular growth control. Pdcd-4 is regulated by phosphorylation-dependent turnover, linking it to PI3K–AKT–mTOR and MAPK signaling inputs that couple extracellular cues to translational reprogramming. Altered PDCD4 expression has been associated with dysregulated proliferation, invasion, and stress responses across multiple cancer-relevant contexts, and it is frequently studied as a node connecting microRNA regulation, apoptosis, and inflammatory signaling. In human cells, PDCD4 perturbation provides a tractable model for dissecting how translational control shapes oncogenic pathways and adaptive responses.

Pdcd-4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PDCD4 expression without altering the underlying DNA sequence.

Pdcd-4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PDCD4 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 PDCD4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pdcd-4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PDCD4 locus and enabling the study of Pdcd-4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pdcd-4 pathway restoration in tumor cells with silenced or reduced PDCD4 expression.

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