Pericentrin 1 CRISPR Activation Plasmid (h): sc-403161-ACT

NUP85 encodes a core component of the Nup107–160 subcomplex of the nuclear pore complex, supporting nucleocytoplasmic transport and maintaining nuclear envelope architecture across the cell cycle. Through its roles in NPC assembly, mitotic progression, and genome stability, NUP85 influences transcriptional programs by regulating the nuclear import/export of signaling mediators and cell-cycle factors. Perturbation of nuclear pore components has been linked to defects in RNA processing, stress responses, and aberrant proliferation in multiple disease contexts, making NUP85 a useful entry point for studying transport-coupled regulation of cell physiology. In human cell models, altered NUP85 activity can be interrogated for effects on nuclear trafficking, checkpoint control, and downstream pathway rewiring.

Pericentrin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUP85 expression without altering the underlying DNA sequence.

Pericentrin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUP85 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 NUP85 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pericentrin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUP85 locus and enabling the study of Pericentrin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pericentrin 1 pathway restoration in tumor cells with silenced or reduced NUP85 expression.

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