Nup88 CRISPR Activation Plasmid (h): sc-404091-ACT

NUP88 encodes Nup88, a core component of the nuclear pore complex that contributes to nucleocytoplasmic transport by organizing the cytoplasmic face of the pore and supporting regulated trafficking of proteins and RNAs. Through interactions with other nucleoporins and transport receptors, Nup88 helps maintain nuclear envelope permeability and influences cell-cycle progression, stress responses, and signal-dependent gene regulation. Altered NUP88 expression or localization has been associated with disrupted transport dynamics and aberrant control of proliferation and differentiation programs. These features make NUP88 a relevant target for mechanistic studies of nuclear transport pathways and their links to genome regulation in human cells.

Nup88 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUP88 expression without altering the underlying DNA sequence.

Nup88 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUP88 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 NUP88 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nup88 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUP88 locus and enabling the study of Nup88-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nup88 pathway restoration in tumor cells with silenced or reduced NUP88 expression.

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