FBXO3 CRISPR Activation Plasmid (m): sc-425412-ACT

Mouse Fbxo3 encodes FBXO3, an F-box substrate receptor within SCF (SKP1–CUL1–F-box) E3 ubiquitin ligase complexes that helps confer specificity to ubiquitin-dependent proteasomal turnover. Through regulated degradation of signaling proteins, FBXO3 contributes to control of inflammatory and stress-responsive pathways, including innate immune signaling and cytokine output. Altered FBXO3 activity has been linked to dysregulated inflammation and immune homeostasis, making it relevant to studies of infection biology, chronic inflammatory phenotypes, and cell-type–specific signaling thresholds. As part of ubiquitin–proteasome system circuitry, Fbxo3 is also useful for probing how protein stability integrates with transcriptional programs and post-translational regulation.

FBXO3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Fbxo3 expression without altering the underlying DNA sequence.

FBXO3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Fbxo3 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 Fbxo3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FBXO3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Fbxo3 locus and enabling the study of FBXO3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FBXO3 pathway restoration in tumor cells with silenced or reduced Fbxo3 expression.

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