Nup205 CRISPR Activation Plasmid (h): sc-404728-ACT

Human NUP205 encodes Nup205, an essential scaffold component of the nuclear pore complex that contributes to nuclear envelope integrity and selective nucleocytoplasmic transport. Nup205 supports assembly and stability of pore architecture and influences multiple processes tied to transport-dependent regulation, including RNA export, protein import, and cell cycle–linked nuclear remodeling. By shaping the efficiency and selectivity of cargo passage, Nup205 can affect transcriptional programs and stress responses that rely on regulated nuclear access. Dysregulation of nuclear pore components, including Nup205-associated pathways, is relevant to studies of proliferative control and cellular phenotypes observed in cancer and neurodevelopmental or neurodegenerative contexts.

Nup205 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUP205 expression without altering the underlying DNA sequence.

Nup205 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUP205 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 NUP205 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nup205 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUP205 locus and enabling the study of Nup205-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nup205 pathway restoration in tumor cells with silenced or reduced NUP205 expression.

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