Nup98 CRISPR Activation Plasmid (h): sc-402182-ACT

Human NUP98 encodes Nup98, a phenylalanine-glycine (FG) repeat nucleoporin that contributes to nuclear pore complex permeability and selective nucleocytoplasmic transport. Beyond its structural role, Nup98 participates in transcriptional regulation, RNA processing, and cell cycle control through interactions with chromatin and transport receptors. NUP98 is frequently implicated in hematologic malignancies via chromosomal rearrangements that generate NUP98 fusion proteins, linking dysregulated gene expression programs to altered differentiation and proliferation. Because nuclear transport and transcriptional control are tightly coupled to stress responses and lineage specification, NUP98 is widely studied in pathways governing nuclear organization and oncogenic transcription networks.

Nup98 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUP98 expression without altering the underlying DNA sequence.

Nup98 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUP98 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 NUP98 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nup98 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUP98 locus and enabling the study of Nup98-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nup98 pathway restoration in tumor cells with silenced or reduced NUP98 expression.

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