Nopp140 CRISPR Activation Plasmid (h): sc-402907-ACT

NOLC1 encodes Nopp140, a nucleolar and Cajal body phosphoprotein that functions as a scaffold for ribonucleoprotein assembly and intranuclear trafficking. Nopp140 participates in ribosome biogenesis by coordinating snoRNP dynamics, pre-rRNA processing, and nucleolar organization, and it supports spliceosomal snRNP maturation through Cajal body interactions. Its activity is regulated by extensive phosphorylation and links to RNA polymerase I–dependent transcriptional programs, nucleolar stress responses, and cell-cycle–coupled growth control. Dysregulation of nucleolar homeostasis and RNA processing pathways involving NOLC1/Nopp140 is relevant to studies of proliferative signaling, genome stability, and stress-adaptive remodeling of gene expression in human cells.

Nopp140 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NOLC1 expression without altering the underlying DNA sequence.

Nopp140 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NOLC1 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 NOLC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nopp140 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NOLC1 locus and enabling the study of Nopp140-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nopp140 pathway restoration in tumor cells with silenced or reduced NOLC1 expression.

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