FBXO30 CRISPR Activation Plasmid (h): sc-402735-ACT

FBXO30 encodes an F-box protein that serves as a substrate recognition component of SCF (SKP1–CUL1–F-box) E3 ubiquitin ligase complexes, coupling target proteins to ubiquitin-dependent proteasomal turnover. Through this role, FBXO30 contributes to regulation of protein homeostasis, cell-cycle progression, and stress-responsive signaling by shaping the stability of key pathway effectors. Altered ubiquitin–proteasome system activity is broadly linked to oncogenic signaling, inflammatory states, and degenerative phenotypes, making FBXO30 expression and function relevant for mechanistic studies in these contexts. In human cell models, modulating FBXO30 provides a handle to interrogate proteostasis networks and downstream transcriptional and phenotypic outputs.

FBXO30 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FBXO30 expression without altering the underlying DNA sequence.

FBXO30 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FBXO30 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 FBXO30 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FBXO30 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FBXO30 locus and enabling the study of FBXO30-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FBXO30 pathway restoration in tumor cells with silenced or reduced FBXO30 expression.

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