FBXO6 CRISPR Activation Plasmid (h2): sc-405919-ACT-2

Human FBXO6 encodes an F-box protein that serves as a substrate recognition component of SCF (SKP1–CUL1–RBX1) E3 ubiquitin ligase complexes, coupling target proteins to ubiquitination and proteasomal turnover. FBXO6 is best characterized in endoplasmic reticulum–associated degradation (ERAD), where it recognizes misfolded glycoproteins via lectin-like interactions and promotes their clearance, thereby influencing proteostasis, ER stress signaling, and quality control of secretory and membrane proteins. Through these functions, FBXO6 intersects with pathways controlling protein folding, ubiquitin-dependent regulation, and cellular stress responses, with reported links to cancer biology, metabolic and inflammatory phenotypes, and disorders associated with aberrant protein homeostasis. Gene editing or perturbation of FBXO6 enables mechanistic studies of substrate specificity within SCF ligases, ERAD dynamics, and the downstream transcriptional and signaling consequences of altered ubiquitin-mediated protein quality control in human cell models.

FBXO6 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous FBXO6 expression without altering the underlying DNA sequence.

FBXO6 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FBXO6 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 FBXO6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FBXO6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FBXO6 locus and enabling the study of FBXO6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FBXO6 pathway restoration in tumor cells with silenced or reduced FBXO6 expression.

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