NOP2 CRISPR Activation Plasmid (h): sc-409656-ACT

Human NOP2 encodes a nucleolar RNA methyltransferase implicated in ribosome biogenesis through modification and maturation of rRNA, supporting assembly of the large ribosomal subunit and efficient protein synthesis. NOP2 activity is tightly linked to nucleolar function, cell-cycle progression, and proliferative growth programs, reflecting its role in coordinating translational capacity with cellular demand. Dysregulated NOP2 expression and nucleolar stress signatures have been reported across multiple tumor contexts, where altered ribosome production and translation control contribute to oncogenic phenotypes. As a result, NOP2 is frequently studied in pathways connecting rRNA processing, chromatin-associated transcriptional regulation, and proteostasis.

NOP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NOP2 expression without altering the underlying DNA sequence.

NOP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NOP2 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 NOP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NOP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NOP2 locus and enabling the study of NOP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NOP2 pathway restoration in tumor cells with silenced or reduced NOP2 expression.

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