NOSIP CRISPR Activation Plasmid (h): sc-404767-ACT

Human NOSIP (nitric oxide synthase interacting protein) is a regulatory adaptor that binds nitric oxide synthases, including eNOS and nNOS, and modulates nitric oxide production and downstream cGMP-dependent signaling. By influencing NOS trafficking, complex formation, and activity, NOSIP can shape cellular processes such as vascular tone regulation, neuronal signaling, and immune-related nitric oxide responses. Dysregulated NOS–NOSIP interactions have been associated with altered redox balance and inflammatory signaling, making NOSIP a useful target for investigating pathways implicated in cardiovascular and neuroinflammatory phenotypes. Studying NOSIP expression and function helps clarify how nitric oxide bioavailability integrates with stress-response and cytokine-linked signaling networks in human cells.

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

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

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