p54/nrb CRISPR Activation Plasmid (m): sc-424729-ACT

Mouse Nono encodes p54/nrb (NONO), a multifunctional nuclear RNA- and DNA-binding protein that localizes to paraspeckles and participates in transcriptional regulation, pre-mRNA splicing, and RNA processing. p54/nrb forms complexes with other DBHS family members to coordinate gene expression programs and can influence DNA damage responses through roles in double-strand break repair and genome stability. Through these activities, NONO contributes to control of cell-cycle progression, stress-adaptive transcription, and nuclear architecture. Dysregulation of NONO-associated RNA regulatory networks has been linked in the literature to altered proliferative signaling and aberrant gene expression states relevant to cancer biology and neurological disease mechanisms.

p54/nrb CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nono expression without altering the underlying DNA sequence.

p54/nrb CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Nono 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 Nono transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p54/nrb expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Nono locus and enabling the study of p54/nrb-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p54/nrb pathway restoration in tumor cells with silenced or reduced Nono expression.

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