Nop56 CRISPR Activation Plasmid (h): sc-404035-ACT

Human NOP56 encodes Nop56, an essential nucleolar core component of box C/D small nucleolar ribonucleoproteins (snoRNPs) that guide 2′-O-methylation and processing of pre-rRNA during ribosome biogenesis. Through its interactions with fibrillarin and other snoRNP factors, Nop56 supports maturation of 18S and 28S rRNA and proper assembly of ribosomal subunits, linking it to global translational capacity and nucleolar homeostasis. Disruption of ribosome biogenesis and nucleolar stress pathways can alter cell-cycle control and proteostasis, making NOP56 function relevant to studies of growth regulation and stress adaptation. NOP56 is also implicated in repeat-expansion–associated neurodegenerative mechanisms in spinocerebellar ataxia type 36 (SCA36), motivating mechanistic work on RNA metabolism and nucleolar dysfunction.

Nop56 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NOP56 expression without altering the underlying DNA sequence.

Nop56 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NOP56 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 NOP56 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nop56 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NOP56 locus and enabling the study of Nop56-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nop56 pathway restoration in tumor cells with silenced or reduced NOP56 expression.

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