C23 (Nucleolin) CRISPR Activation Plasmid (h): sc-400191-ACT

NCL encodes C23 (nucleolin), an abundant nucleolar phosphoprotein that coordinates ribosome biogenesis through rDNA transcription, pre-rRNA processing, and ribosomal subunit assembly. Beyond the nucleolus, nucleolin regulates chromatin organization, DNA damage responses, mRNA stability, and nucleocytoplasmic transport, linking it to cell-cycle progression and cellular stress adaptation. It also participates in signaling and trafficking at the cell surface in some contexts, influencing RNA–protein complex dynamics and translation control. Dysregulated nucleolin expression or localization is frequently associated with proliferative phenotypes, genome instability, and altered RNA metabolism, making it relevant to studies of cancer biology and neurodegenerative stress pathways.

C23 (Nucleolin) CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCL expression without altering the underlying DNA sequence.

C23 (Nucleolin) CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCL 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 NCL transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous C23 (Nucleolin) expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCL locus and enabling the study of C23 (Nucleolin)-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of C23 (Nucleolin) pathway restoration in tumor cells with silenced or reduced NCL expression.

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