Ran CRISPR Activation Plasmid (h): sc-417223-ACT

Human RAN encodes Ran, a small Ras-related GTPase that establishes the RanGTP/RanGDP gradient required for nucleocytoplasmic transport through the nuclear pore complex. Ran regulates importin/exportin-mediated cargo trafficking, mitotic spindle assembly, nuclear envelope reformation, and RNA processing, integrating with cell cycle control and checkpoint signaling. Because these processes govern genome stability and global gene expression programs, altered Ran activity has been associated with proliferative phenotypes and dysregulated nuclear transport observed across multiple disease contexts, including cancer and neurodegeneration. RAN is therefore widely studied as a node connecting nuclear transport, mitosis, and stress-adaptive transcriptional responses.

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

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

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