Nup155 CRISPR Activation Plasmid (h): sc-405347-ACT

NUP155 encodes Nup155, a core component of the nuclear pore complex that contributes to nuclear envelope integrity and the selective transport of proteins and RNAs between the nucleus and cytoplasm. Nup155 participates in nucleocytoplasmic trafficking, NPC assembly, and cell cycle–linked remodeling of the nuclear envelope, processes that influence gene regulation and cellular homeostasis. Disruption of nuclear pore function can perturb RNA processing and signaling networks that rely on regulated nuclear import/export, and NUP155 has been linked to human disease contexts involving nuclear transport defects, including cardiac conduction phenotypes and broader nuclear envelope–associated pathobiology. These features make NUP155 a useful target for studying how NPC architecture shapes transcriptional programs and stress responses.

Nup155 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUP155 expression without altering the underlying DNA sequence.

Nup155 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUP155 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 NUP155 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nup155 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUP155 locus and enabling the study of Nup155-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nup155 pathway restoration in tumor cells with silenced or reduced NUP155 expression.

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