Nischarin CRISPR Activation Plasmid (h): sc-404823-ACT

Human NISCH encodes Nischarin, a cytosolic scaffolding protein originally identified as an imidazoline receptor–interacting factor that modulates integrin α5 signaling and cytoskeletal organization. Nischarin coordinates cell adhesion, migration, and endocytic trafficking through interactions with focal adhesion and Rho-family GTPase pathways, influencing actin remodeling and directional motility. It has been linked to regulation of intracellular signaling nodes such as MAPK cascades and vesicle transport programs that shape cellular polarity and invasive behavior. Altered NISCH expression has been reported across multiple disease contexts, including cancer biology and neurobehavioral phenotypes, making it relevant for mechanistic studies of migration, metastasis-associated pathways, and neuronal signaling.

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

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

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