CUL-3 CRISPR Activation Plasmid (h): sc-416925-ACT

Human CUL3 encodes cullin-3 (CUL-3), a core scaffold of CRL3 E3 ubiquitin ligase complexes that couple BTB-domain substrate adaptors to RBX1 for ubiquitin transfer and proteasomal degradation. Through selective turnover of regulatory proteins, CUL-3 influences proteostasis, cell-cycle progression, oxidative stress responses, and cytoskeletal organization, including pathways linked to KEAP1–NRF2 signaling. Dysregulated CRL3 activity can perturb redox homeostasis and checkpoint control, making CUL3 a frequently studied node in cancer biology and stress-adaptation programs. Alterations in CUL3-dependent ubiquitination networks are also used to model mechanisms relevant to neurodevelopmental and cardiovascular phenotypes in cellular systems.

CUL-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CUL3 expression without altering the underlying DNA sequence.

CUL-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CUL3 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 CUL3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CUL-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CUL3 locus and enabling the study of CUL-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CUL-3 pathway restoration in tumor cells with silenced or reduced CUL3 expression.

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