prefoldin 4 CRISPR Activation Plasmid (h): sc-416619-ACT

PFDN4 encodes prefoldin 4, a subunit of the prefoldin co-chaperone complex that captures nascent or unfolded polypeptides and delivers them to the TRiC/CCT chaperonin for productive folding. This activity supports cytoskeletal proteostasis by facilitating the maturation of actin and tubulin and helps maintain protein homeostasis under cellular stress. Through its role in chaperone-mediated folding, PFDN4 contributes to pathways governing cell cycle progression, proteostasis networks, and stress-response signaling. Dysregulated chaperone function and altered prefoldin complex activity have been linked to proteostasis imbalance observed in cancer and neurodegeneration research contexts.

prefoldin 4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PFDN4 expression without altering the underlying DNA sequence.

prefoldin 4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PFDN4 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 PFDN4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous prefoldin 4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PFDN4 locus and enabling the study of prefoldin 4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of prefoldin 4 pathway restoration in tumor cells with silenced or reduced PFDN4 expression.

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