Ran BP-M CRISPR Activation Plasmid (h): sc-403548-ACT

Human RANBP9 encodes Ran BP-M, a scaffolding protein that associates with the Ran GTPase system and contributes to nucleocytoplasmic transport, signal integration, and macromolecular complex assembly. Ran BP-M has been linked to regulation of cytoskeletal organization and cell adhesion through interactions with receptor-associated and kinase signaling modules, supporting context-dependent control of proliferation and stress responses. Altered RANBP9 expression and network connectivity have been observed in studies of cancer biology and neurodegeneration-related pathways, where it can influence transcriptional programs and protein stability. These properties make RANBP9 a useful node for dissecting pathway crosstalk between nuclear transport machinery, signaling cascades, and proteostasis.

Ran BP-M CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RANBP9 expression without altering the underlying DNA sequence.

Ran BP-M CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RANBP9 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 RANBP9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ran BP-M expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RANBP9 locus and enabling the study of Ran BP-M-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ran BP-M pathway restoration in tumor cells with silenced or reduced RANBP9 expression.

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