Exportin 5 CRISPR Activation Plasmid (h): sc-403402-ACT

XPO5 encodes Exportin 5, a RanGTP-dependent nuclear export receptor that shuttles pre-miRNAs and select structured RNAs from the nucleus to the cytoplasm, enabling subsequent processing by DICER and assembly of RNA-induced silencing complexes. By controlling the supply of pre-miRNAs to the cytoplasmic miRNA biogenesis pathway, Exportin 5 influences post-transcriptional gene regulation, cellular differentiation programs, stress responses, and proliferation. Perturbation of XPO5 expression or transport activity can reshape global miRNA profiles and downstream transcriptional networks, making it relevant to studies of oncogenic signaling, metastasis-associated phenotypes, and genome stability. Exportin 5 also interfaces with broader nucleocytoplasmic transport processes coordinated by the Ran cycle, linking RNA trafficking to nuclear organization and gene expression homeostasis.

Exportin 5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous XPO5 expression without altering the underlying DNA sequence.

Exportin 5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the XPO5 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 XPO5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Exportin 5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native XPO5 locus and enabling the study of Exportin 5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Exportin 5 pathway restoration in tumor cells with silenced or reduced XPO5 expression.

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