Skp2 p45 CRISPR Activation Plasmid (h): sc-400534-ACT

SKP2 encodes the F-box protein Skp2 p45, a substrate-recognition component of the SCF (SKP1–CUL1–F-box) E3 ubiquitin ligase complex that controls proteasomal turnover of key cell-cycle regulators. By promoting ubiquitination of targets such as CDKN1B/p27^Kip1 and other growth-inhibitory proteins, Skp2 supports G1/S transition, DNA replication competence, and coordinated checkpoint responses. SKP2 activity interfaces with PI3K–AKT, RB–E2F, and TGF-β-linked signaling programs that shape proliferation, senescence, and stress adaptation. Dysregulated SKP2 expression or function has been associated with altered cell-cycle control and genomic instability in multiple disease-relevant contexts, making it a common node for mechanistic studies of oncogenic signaling and protein homeostasis.

Skp2 p45 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SKP2 expression without altering the underlying DNA sequence.

Skp2 p45 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SKP2 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 SKP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Skp2 p45 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SKP2 locus and enabling the study of Skp2 p45-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Skp2 p45 pathway restoration in tumor cells with silenced or reduced SKP2 expression.

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