Nup133 CRISPR Activation Plasmid (m): sc-433405-ACT

Mouse Nup133 encodes a core structural component of the nuclear pore complex (NPC) Nup107–160 subcomplex, which scaffolds nuclear pore assembly and supports nucleocytoplasmic transport. By helping organize NPC architecture, Nup133 influences RNA and protein trafficking, cell-cycle progression, and nuclear envelope dynamics, including reassembly after mitosis. Perturbation of NPC composition can alter global gene expression programs through disrupted transport of transcription factors and RNA-processing machinery. As a central nucleoporin, Nup133 is frequently studied in contexts linking nuclear transport and nuclear envelope integrity to developmental phenotypes and cellular stress responses.

Nup133 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nup133 expression without altering the underlying DNA sequence.

Nup133 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Nup133 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 Nup133 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nup133 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Nup133 locus and enabling the study of Nup133-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nup133 pathway restoration in tumor cells with silenced or reduced Nup133 expression.

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