p54/nrb Lentiviral Activation Particles (m): sc-424729-LAC

Mouse Nono encodes p54/nrb (NONO), a multifunctional nuclear RNA- and DNA-binding protein of the DBHS family that forms heterodimers with SFPQ/PSPC1 to organize paraspeckles and coordinate transcription, pre-mRNA splicing, and RNA retention. p54/nrb participates in DNA damage response and repair, including roles in non-homologous end joining, and helps couple signaling-dependent transcription to RNA processing through interactions with RNA polymerase II–associated complexes. Through regulation of gene expression programs controlling cell cycle progression, differentiation, and stress responses, altered NONO activity has been linked to genome instability and aberrant transcriptional networks relevant to cancer and neurodevelopmental phenotypes. As a scaffold for ribonucleoprotein assemblies, p54/nrb also influences nuclear body dynamics and long noncoding RNA–mediated regulation, making it a useful target for mechanistic studies of nuclear organization.

p54/nrb Lentiviral Activation Particles (m) address this need by packaging the complete synergistic activation mediator (SAM) transcriptional activation system into transduction-ready, high-titer lentiviral particles, enabling efficient Nono upregulation across a broader range of human cell types.

p54/nrb Lentiviral Activation Particles (m) deliver all functional components of the synergistic activation mediator (SAM) system via lentiviral transduction. The system comprises three particle preparations co-transduced into target cells: one encoding catalytically inactive dCas9 (D10A and N863A mutations) fused to the VP64 transactivation domain with a blasticidin resistance gene; one encoding the MS2-p65-HSF1 fusion protein with a hygromycin resistance gene; and one encoding a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers with a puromycin resistance gene. Following lentiviral transduction and genomic integration of the expression cassettes, the SAM components are stably expressed and assemble at the target locus within the proximal promoter region upstream of the Nono transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous p54/nrb expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native Nono genomic locus and regulatory architecture.

The lentiviral format offers several practical advantages: stable genomic integration supports heritable activation across cell divisions; high-titer particle preparations eliminate the need for in-house viral production; and compatibility with primary, non-dividing, and transfection-resistant cell types expands experimental accessibility. Successful transduction can be confirmed and enriched through triple antibiotic selection using puromycin, hygromycin, and blasticidin.

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