NAB2 CRISPR Activation Plasmid (h): sc-402117-ACT

NAB2 (NGFI-A binding protein 2) is a nuclear transcriptional coregulator that modulates immediate early gene responses by interacting with EGR family transcription factors, shaping stimulus-dependent programs controlling differentiation, proliferation, and cellular stress adaptation. By influencing transcriptional outputs downstream of MAPK/ERK-linked signaling and other activity-regulated pathways, NAB2 helps tune gene networks involved in growth control and lineage specification. Altered NAB2 regulation is relevant to aberrant transcriptional states observed in cancer biology and other proliferative disorders, and NAB2 gene fusions and dysregulated expression have been associated with tumorigenic processes in specific contexts. These properties make NAB2 a useful target for dissecting transcription factor–coregulator dynamics and mapping stimulus-responsive gene expression circuits in human cells.

NAB2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NAB2 expression without altering the underlying DNA sequence.

NAB2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NAB2 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 NAB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NAB2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NAB2 locus and enabling the study of NAB2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NAB2 pathway restoration in tumor cells with silenced or reduced NAB2 expression.

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