FBXO10 CRISPR Activation Plasmid (h): sc-406159-ACT

FBXO10 encodes an F-box protein that functions as a substrate-recognition component of SCF (SKP1–CUL1–F-box) E3 ubiquitin ligase complexes, directing specific proteins for ubiquitination and proteasome-dependent turnover. Through regulated protein degradation, FBXO10 contributes to control of signaling amplitude, cell-cycle and stress-response programs, and maintenance of proteostasis. Dysregulated SCF-mediated ubiquitin signaling and altered FBXO10 activity have been linked to aberrant survival signaling and lymphoid biology, supporting its relevance in studies of apoptosis, oncogenic pathway buffering, and immune cell homeostasis. FBXO10 is therefore of interest for dissecting ubiquitin–proteasome system wiring and context-dependent regulation of key pathway nodes.

FBXO10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FBXO10 expression without altering the underlying DNA sequence.

FBXO10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FBXO10 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 FBXO10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FBXO10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FBXO10 locus and enabling the study of FBXO10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FBXO10 pathway restoration in tumor cells with silenced or reduced FBXO10 expression.

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