N4BP2 CRISPR Activation Plasmid (h): sc-412585-ACT

N4BP2 (NEDD4 binding protein 2) is a large intracellular protein implicated in ubiquitin-dependent signaling through its reported interaction with NEDD4 family E3 ligases, linking it to regulation of protein turnover and signal transduction. It has been associated with control of cytoskeletal organization and cell motility, consistent with roles in adhesion dynamics and intracellular trafficking. N4BP2 expression and genetic alteration patterns have been explored in cancer and immune-related contexts, supporting its relevance to pathways that shape proliferation, stress responses, and inflammatory signaling. These features make N4BP2 a useful target for dissecting ubiquitin pathway crosstalk and downstream transcriptional programs in human cell models.

N4BP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous N4BP2 expression without altering the underlying DNA sequence.

N4BP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the N4BP2 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 N4BP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous N4BP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native N4BP2 locus and enabling the study of N4BP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of N4BP2 pathway restoration in tumor cells with silenced or reduced N4BP2 expression.

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