Ran BP-3 CRISPR Activation Plasmid (h): sc-405001-ACT

RANBP3 encodes Ran BP-3, a RanGTP-binding protein that functions as a cofactor for CRM1/XPO1-dependent nuclear export, helping regulate the nucleocytoplasmic distribution of proteins and RNAs. By modulating RanGTP gradients and export complex assembly, Ran BP-3 contributes to control of gene expression programs, cell cycle progression, and stress-responsive signaling through nuclear transport dynamics. Dysregulated nuclear export is a recurrent feature of cancer and other proliferative or inflammatory states, making RANBP3 a useful target for mechanistic studies linking transport defects to altered transcriptional networks. In human cells, perturbing RANBP3 expression is commonly leveraged to probe pathway connectivity between nuclear trafficking, transcriptional regulation, and proteostasis.

Ran BP-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RANBP3 expression without altering the underlying DNA sequence.

Ran BP-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RANBP3 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 RANBP3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ran BP-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RANBP3 locus and enabling the study of Ran BP-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ran BP-3 pathway restoration in tumor cells with silenced or reduced RANBP3 expression.

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