Nup37 CRISPR Activation Plasmid (h): sc-409458-ACT

NUP37 encodes Nup37, a core subunit of the Nup107–160 complex that scaffolds nuclear pore complex (NPC) assembly and supports selective nucleocytoplasmic transport. By contributing to NPC biogenesis and permeability, Nup37 influences RNA and protein trafficking, cell-cycle progression, and genome regulatory programs that depend on regulated nuclear import/export. Altered expression or perturbation of nucleoporins is associated with defects in mitotic fidelity, transcriptional control, and stress responses, linking NPC dysfunction to proliferative and developmental phenotypes. NUP37 is therefore relevant for studying how nuclear transport architecture interfaces with chromatin regulation and pathways frequently dysregulated in cancer-related cell states.

Nup37 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUP37 expression without altering the underlying DNA sequence.

Nup37 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUP37 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 NUP37 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nup37 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUP37 locus and enabling the study of Nup37-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nup37 pathway restoration in tumor cells with silenced or reduced NUP37 expression.

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