Emi2 CRISPR Activation Plasmid (h): sc-405054-ACT

FBXO43 encodes the human protein Emi2, an F-box–containing regulator best known for controlling cell-cycle transitions through modulation of APC/C activity and downstream turnover of mitotic regulators. By restraining anaphase-promoting complex function until appropriate cell-cycle cues are met, Emi2 contributes to coordinated progression through M phase and maintenance of genomic integrity. Altered expression or regulation of cell-cycle ubiquitin–proteasome pathways involving F-box proteins is frequently linked to proliferative phenotypes and chromosomal instability observed in cancer biology. FBXO43/Emi2 therefore serves as a mechanistic entry point for studying ubiquitin-mediated proteolysis, checkpoint control, and deregulated proliferation in human cellular models.

Emi2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FBXO43 expression without altering the underlying DNA sequence.

Emi2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FBXO43 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 FBXO43 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Emi2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FBXO43 locus and enabling the study of Emi2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Emi2 pathway restoration in tumor cells with silenced or reduced FBXO43 expression.

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