CUL-1 CRISPR Activation Plasmid (h): sc-400972-ACT

Human CUL1 encodes cullin-1 (CUL-1), a core scaffold of SCF (SKP1–CUL1–F-box) E3 ubiquitin ligase complexes that coordinate substrate recognition and ubiquitin transfer to regulate proteasomal turnover. By controlling the stability of key cell-cycle and signaling regulators such as cyclins, CDK inhibitors, and components of NF-κB and Wnt pathways, CUL-1 helps maintain orderly progression through G1/S transitions and signal-dependent transcriptional programs. CUL-1 activity is modulated by neddylation/deneddylation cycles, linking it to dynamic regulation of ubiquitin-mediated proteostasis. Dysregulation of SCF/CUL1-dependent ubiquitination has been associated with aberrant proliferation, genomic instability, and altered inflammatory signaling in multiple disease-relevant contexts.

CUL-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CUL1 expression without altering the underlying DNA sequence.

CUL-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CUL1 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 CUL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CUL-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CUL1 locus and enabling the study of CUL-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CUL-1 pathway restoration in tumor cells with silenced or reduced CUL1 expression.

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